Excitation Transfer in Phycobiliproteins

Encyclopedia of Plant Physiology

New Series Volume 19

Editors A. Pirson, Göttingen M. H. Zimmermann, Harvard

Photosynthesis IQ Photosynthetic Membranes and Light Harvesting Systems

Edited by

L. A. Staehelin and C. J. Arntzen

Contributors J.M.Anderson B.Andersson C.J.Arntzen R.Bachofen J.Barber J.Barrett R.E.Blankenship RBöger J.Breton T.M.Bricker R.J.Cogdell J. Deisenhofer R.A.Dilley B.A.Diner T.J.Donohue P.L.Dutton R.C.Fuller E.Gantt N. E. Geacintov J. A. Guikema W.Haehnel G.Hauska J.E.Hearst A.J.Hoff D.Holten A.R.Holzwarth A.Joliot P.Joliot S.Kaplan K.Keegstra R.S.Knox D.J.Kyle C.Lütz P.Mathis R.E.McCarty B.A.Melandri H.Michel D.J.Murphy C.M.Nalin J.R.Norris I.Ohad D.R.Ort H. B.Pakrasi W.W.Parson R.C.Prince A.Rawyler I. J.Ryrie G.Sandmann K.Sauer H.Scheer P.Sétif L.A.Sherman P.A.Siegenthaler D.J.Simpson S.G.Sprague L. A. Staehelin H.Strotmann J.P.Thornber D.M.Tiede G. Van Brakel A. R. Varga G. Venturoli J. Whitmarsh V.Wiemken F.-A.Wollman C.F.Yocum H. Zuber

Springer-Verlag Berlin Heidelberg New York Tokyo Professor L. ANDREW STAEHELIN Department of Molecular, Cellular, and Developmental Biology University of Colorado Campus Box 347 Boulder, CO 80309 USA

Professor CHARLES J. ARNTZEN E.I. du Pont de Nemours & Co. Central Research and Development Department Experimental Station Bldg. 402, Room 3226 Wilmington, DE 19898 USA

With 163 Figures

ISBN 3-540-16140-6 Springer-Verlag Berlin Heidelberg New York Tokyo ISBN 0-387-16140-6 Springer-Verlag New York Heidelberg Berlin Tokyo

Library of Congress Cataloging-in-Publication Data. (Revised for vol. 3) Main entry under title: Photosyn• thesis. (Encyclopedia of plant physiology; new ser., v. 5-6, 19) Includes bibliographies and indexes. Contents: 1. Photosynthetic electron transport and photophosphorylation, edited by A. Trebst and M. Avron. 2. Photosynthetic carbon metabolism and related processes, edited by M. Gibbs and E. Latzko. 3. Photosyn• thetic membranes and light harvesting systems, edited by L.A. Staehelin and C.J. Arntzen. 1. Photosynthesis - Collected works. I. Trebst, Achim. II. Alberte, R.S. III. Series. QK711.2.E5 vol. 5-6 581.1s [581.1'3342] 76-30357 [QK882] ISBN 0-387-07962-9 (v. 1) This work is subjected to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically those of translation, reprinting, re-use of illustrations, broadcasting, reproduction by photocopying machine or similar means, and storage in data banks. Under § 54 of the German Copyright Law where copies are made for other than private use, a fee is payable to „Verwertungsgesellschaft Wort" Munich © by Springer-Verlag Berlin Heidelberg 1986 Printed in Germany The use of registered names, trademarks, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. Typesetting, printing and bookbinding: Universitätsdruckerei H Stürtz AG, Würzburg 2131/3130-543210 Preface

The Encyclopedia of Plant Physiology series has turned several times to the topic of photosynthesis. In the original series, two volumes edited by A. PERSON and published in 1960 provided a broad overview of the entire field. Although the New Series has devoted three volumes to the same topic, the overall breadth of the coverage has had to be restricted to allow for greater in-depth treatment of three major areas of modern photosynthesis research: I. Photosynthetic Elec• tron Transport and Photophosphorylation (Volume 5 edited by A. TREBST and M. AVRON, and published in 1977); II. Photosynthetic Carbon Metabolism and Related Processes (Volume 6 edited by M. GIBBS and E. LATZKO, and published in 1979); and III. Photosynthetic Membranes and Light-Harvesting Systems (this volume). As we approached the organization of the current volume, we chose a set of topics for coverage that would complement the earlier volumes, as well as provide updates of areas that have seen major advances in recent years. In addition, we wanted to emphasize the following changes in the study of photo• synthetic systems which have become increasingly important since 1977: the trend toward increased integration of biochemical and biophysical approaches to study photosynthetic membranes and light-harvesting systems, and a renewed appreciation of the structural parameters of membrane organization. Due to the increased complexity of the field, we also decided to try a new format for our volume to better serve the following two purposes. First, we believe a review volume on photosynthetic membranes should serve as a refer• ence source for nonspecialists interested in obtaining an overview of both oxy• genic and anoxygenic photosynthesis. This need has been answered by the inclu• sion of five introductory chapters which summarize the main broad topic areas of the volume. We also recognize that a review volume should provide insight to the "state of the art" in specific research areas which have seen major recent advances. To this end, Chapters 6 through 11 have been organized such that each consists of a number of minireviews related to a common theme. All of the 43 minireviews are authored by highly regarded specialists, and focus on recent research highlights and interpretations of significant new findings. Great emphasis has been placed on the integration of the materials covered in the introductory chapters and in the minireviews. Extensive cross-referencing has been used to allow easy transitions by the reader from a general to a specialized coverage of a topic. Similarly, all minireviews contain references to the appropriate introductory chapters, as well as to other minireviews. VI Preface

With students in mind, the authors of the introductory chapters have stressed integrative and comparative aspects of their topics. This type of approach is becoming more and more relevant in photosynthesis research, thanks to the convergence of information coming from structural, biophysical, and biochemi• cal studies. Indeed, it is truly exciting to witness the progress being made toward the goal of a molecular understanding of the diverse biophysical and biochemical reactions associated with photosynthetic membranes and light-harvesting sys• tems. The specialized chapters in this volume begin with the topic of light harvest• ing by photosynthetic membranes. The minireviews of Chapter 6 summarize biochemical and structural studies of light-harvesting assemblages, with empha• sis on the light-harvesting components of bacteria and algae, since higher plant chloroplast components are extensively reviewed in Chapter 3. Whereas Chapter 6 emphasizes the biochemical diversity in light-harvesting systems, Chapter 7 consists of minireviews which discuss unifying concepts governing light-harvest• ing events. All authors in this section are concerned with photon absorption and structural parameters of the pigment bed that determine the efficiency of excitation energy transfer to reaction centers. The most fundamental result of photosynthetic light reactions is the conver• sion of excitation energy, derived from absorbed light, into stable chemical form. This occurs in the reaction center (RC). In the last 5 years there have been major advancements in the understanding of these processes - especially by those groups who have focused their work on photosynthetic bacteria. The identity of the cofactors (chlorophyll, pheophytin, quinones, etc.) involved in the initial charge separations and the events involved in charge stabilization are now highly defined. The minireviews of Chapter 8 present various aspects of this rapidly moving field, ranging from energetic considerations of the RC to discussions of similarities and differences among the different types of protein which comprise different reaction centers. Chapter 9 makes a transition from the highly defined bacterial reaction centers into the less well understood photosystems I and II of green plants. The inclusion of several minireviews devoted to the complexity of reactions in photosystem II, for example, reflects the wide diversity in studies of a system capable of extracting electrons from water and catalyzing a stable charge separa• tion that results in reduction of the plastoquinone pool. These studies extend from detailed understanding of the primary reactions to physiological adapta• tion of the process to light and chemical (herbicide) stresses. The reaction centers of prokaryotic and eukaryotic photosynthetic mem• branes produce high energy electrons which are utilized in electron transport reactions. The energy released in these reactions is coupled to ATP synthesis. The membrane components and processes involved in the energy-coupling reac• tions are the topic of Chapter 10. Minireviews in this section are strongly bio• chemical in emphasis, with special reference to the structural organization of membranes and the protein complexes which mediate proton translocation and ATP biosynthesis. Specific enzymes involved in electron transport and inhibitors which affect them are reviewed in light of information obtained since 1977. Preface VII

The last chapter of this volume (Chap. 11) deals with the use of integrative approaches to study processes associated with control of photosynthetic mem• brane assembly and maintenance. Chapter 11 includes subject material ranging from comparative structural analysis of photosynthetic membranes (develop• mental diversity) to the use of physical analysis of membranes or simulated membrane systems to characterize functional components. The minireviews of this chapter will be of increasing value as the field of membrane biosynthesis and assembly matures to use more information about physical and biochemical features of the photosynthetic membranes. We can anticipate the advent of use of genetic engineering tools to manipulate photosynthetic membranes, and the rapid expansion of knowledge in this area. In summary, this volume is a selection of both overview chapters and numer• ous topical speciality reviews. It should be useful as a reference source and as a teaching aid for individuals interested in the rapidly expanding field of photosynthetic membranes.

Boulder and Wilmington, Spring 1986 L.A. STAEHELIN C.J. ARNTZEN

Contents

1 Chloroplast Structure and Supramolecular Organization of Photosynthetic Membranes L.A. STAEHELIN (With 45 Figures) 1 Introduction 1 2 Chloroplast Morphology 1 2.1 Light Microscopic Observations 1 2.2 The Chloroplast Envelope 2 2.2.1 Higher Plants 2 2.2.1.1 Morphology and Isolation 2 2.2.1.2 Infoldings of the Inner Envelope Membrane 4 2.2.2 Algae 5 2.3 Chloroplast Stroma 10 2.3.1 Matrix Components 10 2.3.2 Plastoglobuli 11 2.3.3 Pyrenoids 11 2.3.4 Eyespots 13 2.4 Arrangement of Thylakoids 14 2.4.1 Thylakoids of Oxygen-Evolving Prokaryotes 16 2.4.1.1 Cyanobacteria 16 2.4.1.2 Prochlorons 19 2.4.2 Higher Plant Thylakoids 19 2.4.2.1 Three-Dimensional Organization of Grana and Stroma Thylakoids 19 2.4.2.2 Normal Development of Grana and Stroma Thylakoids 20 2.4.2.3 Thylakoid Membrane Nomenclature 22 2.4.2.4 Naturally and Experimentally Induced Variations in the Ratio of Stacked to Unstacked Membrane Regions . . 23 2.4.3 Green Algal Thylakoids 25 2.4.4 Euglenoid Thylakoids 25 2.4.5 Red Algal Thylakoids 26 2.4.6 Chromophytan Thylakoids 28 2.4.6.1 Cryptophyceae (Cryptomonads) 28 2.4.6.2 Phaeophyceae (Brown Algae) and Bacillariophyceae (Diatoms) 28 2.4.6.3 Chrysophyceae (Golden-Brown-Algae) and Xanthophyceae (Yellow-Green Algae) 29 2.4.6.4 Dinophyceae (Dinoflagellates) 29 3 Supramolecular Organization of Thylakoids 30 3.1 Freeze-Fracture (-Etch) Electron Microscopy 30 3.2 Higher Plant and Green Algal Thylakoids 32 3.2.1 Photosystem II Complexes 34 3.2.2 Photosystem I Complexes 39

3.2.3 Cytochrome b6/f Complex 40 3.2.4 CFQ-CF! ATP Synthase 42 3.2.5 Chlorophyll a/b Light-Harvesting Complex 44 X Contents

3.2.5.1 "Mobile" Versus "Bound" Forms of Chi a/b LHC . . 44 3.2.5.2 Chi a/b LHC as Membrane Adhesion Factor 45 3.2.5.3 Reconstitution and Structural Characterization .... 45 3.2.6 Why Do Chloroplasts Need Grana? 47 3.2.7 Supramolecular Organization of Thylakoids - a Summary ... 48 3.3 Prochloron Thylakoids 51 3.4 Thylakoid Membranes of Cyanobacteria, Red Algae and Cyanelles 53 3.4.1 Identification of Freeze-Fracture Particles 53 3.4.2 Relationship Between Phycobilisomes and EF Particles (PS II Complexes) 56 3.5 Thylakoid Membranes of Chromophyta 58 3.5.1 Cryptophyceae 59 3.5.2 Phaeophyceae and Dinophyceae 60 4 Experimental Modification of Thylakoid Membrane Structure 61 4.1. Structural Changes Associated with in Vitro Unstacking and Restacking of Thylakoids 61 4.2 State I/State II Transitions - Functional Regulation Mediated by Structural Rearrangements 64 4.3 Temperature-Induced Changes in Thylakoid Membrane Organization 66 4.3.1 Effects of Heat Stress 66 4.3.2 Chilling Effects 67 4.3.2.1 Higher Plants 67 4.3.2.2 Thermophilic Cyanobacteria 69 4.4 Prothylakoids and Prolamellar Bodies 70 References 72

2 Photosynthetic Light Reactions - Physical Aspects K. SAUER (With 5 Figures) 1 Introduction 85 2 Radiation - Sunlight 86 3 The Excitation of Pigment Molecules 88 4 Excitation Transfer 90 5 Electron Transfer - Charge Separation in the Reaction Center 92 6 The Reaction Center - Structure and Function 93 7 Conclusion 95 References 96

3 Biochemical Characterization and Structure of Pigment-Proteins of Photosynthetic Organism J.P. THORNBER (With 4 Figures) 1 Introduction 98 2 Development of the Subject 100 2.1 Historical Development of Our Understanding of Plant Chlorophyll- Proteins 100 2.2 Development of Our Knowledge of Pigment-Proteins of Photosynthetic Bacteria 106 3 The State of Knowledge of the Biochemical Organization of Photosynthetic Pigments in Plants and Bacteria 107 3.1 Photosystem I of Green Plants 109 3.1.1 Core Complex I (CC I) 109 3.1.2 Light-Harvesting Complex I (LHC I) Ill 3.2 Photosystem II 112 3.2.1 CCU 112 3.2.2 LHC II 113 Contents XI

3.3 Pigment-Protein Complexes of the Algae 116 3.4 Pigment-Protein Complexes of the Cyanobacteria 117 3.5 Purple Bacteria 119 3.5.1 Photochemical Reaction Centers 119 3.5.2 The Light-Harvesting Pigment-Protein Complexes 123 3.5.2.1 The B-800-850 Complex 123 3.5.2.2 The B-890-Protein Complex 125 3.5.3 Topography of the Antenna and Reaction Center Complexes 125 3.6 Chlorophyll-Protein Complexes of the Green Bacteria 126 4 Concluding Remarks 127 4.1 Isolation of the Pigment-Protein Complexes 128 4.2 The Structure of the Chlorophyll-Protein Complexes 132 References 135

4 Energy Transduction in Oxygenic Photosynthesis: an Overview of Structure and Mechanism D.R. ORT (With 7 Figures) 1 Introduction 143 2 The Chloroplast Thylakoid Membrane 146 2.1 Thylakoid Membrane Morphology 146 2.2 Chemical Composition and Physical Properties of the Thylakoid Membrane 149 2.2.1 Composition 149 2.2.1.1 Lipids 150 2.2.1.2 Polypeptides 152 2.2.1.3 Carbohydrates 154 2.2.2 Physical Properties 154 2.3 Distribution of Thylakoid Membrane Components 156 2.3.1 Transmembrane Asymmetry 156 2.3.2 Lateral Asymmetry 157 3 Photosystem II Complex 158 3.1 Composition and Structure 158 3.2 Primary Electron Transfer Events 161 3.3 The Acceptor-Side Reactions of Photosystem II Electron Transfer . . 162 3.3.1 The Two-Electron Gate 163 3.3.2 Mode of Inhibition by Photosystem II Herbicides 164 3.4 Water Oxidation and Donor-Side Reactions 165 3.4.1 The Reduction of P + -680 165 3.4.2 Water Oxidation 166 4 Photosystem I Complex 168 4.1 Composition and Structure 168 4.2 Primary Electron Transfer Events 169 4.3 Electron Donors to Photosystem I 170 4.4 The Acceptor-Side Components and Reactions of Photosystem I . . 171 5 Intersystem Electron Transfer 172

5.1 Cytochrome b6/f Complex 173 5.1.1 Composition and Structure 173 5.1.2 Mechanism of the Oxidation of Plastoquinol 174

5.1.3 Involvement of the Cytochrome b6/f Complex in Cyclic Electron Flow Around Photosystem I 176 5.2 Mobile Electron Carriers 177 6 Photophosphorylation 179 6.1 Mechanism and Energetic Considerations 179 6.2 "Phosphorylating Unit" 181 6.2.1 Distribution and Stoichiometry of Components of the Phosphorylating Unit 181 XII Contents

6.2.2 Localized vs. Delocalized Coupling 182 6.3 Coupling Factor Complex 184 6.3.1 Structure 184 6.3.2 Regulation of Catalytic Activity 185 6.3.3 Catalytic Mechanisms 186 7 Concluding Remarks 188 References 189

5 Energy Transduction in Anoxygenic Photosynthesis P.L.DUTTON (With 1 Figure) 1 Introduction 197 2 The Photosynthetic Bacterial Membrane 201 3 Membrane-Associated Energy-Transducing Enzymes 202 3.1 The Light-Harvesting System 203 3.2 The Reaction Center 203 3.3 The Oxido-Reductases 206 3.3.1 Direct NAD(P)+Reduction 206 3.3.2 Direct Quinone Reduction 207 3.3.3 The be! Complexes 208 + 210 3.3.4 NAD Reduction Coupled to AjiH* 3.3.5 Direct Versus Indirect NAD(P)+ Reduction 211 3.3.6 The Supply of Electrons from Oxidizable Substrates 212 3.4 The Reaction Center - Cytochrome c Complex 220 3.4.1 General Observations 221 3.4.2 The Rps. viridis Structure 224 3.4.3 Further Notes on Conformations, States, and Reaction Rates 227 3.5 Membrane Translocators 228 3.6 The ATP-Synthase 229 4 Concluding Remarks 230 References 232

6 Comparative Biochemistry of Light-Harvesting Systems

6.1 Primary Structure and Function of the Light-Harvesting Polypeptides from Cyanobacteria, Red Algae, and Purple Photosynthetic Bacteria H. ZUBER (With 4 Figures) 1 Introduction 238 2 The Polypeptides of the Extra-Membrane Light-Harvesting Pigment- Protein Complexes (Phycobiliproteins) of Cyanobacteria and Red Algae 241 2.1 Structural Organization of the Phycobiliproteins 241 2.2 Primary Structures of the Phycobiliproteins: Structural and Functional Implications 242 3 The Polypeptides of the Intra-Membrane Light-Harvesting Pigment- Protein Complexes of Purple Photosynthetic Bacteria 245 3.1 Types and Molecular Organization of the Antenna Complexes . . . 245 3.2 Primary Structures and the Structural and Functional Role of the Light-Harvesting Polypeptides 246 4 Concluding Remarks 250 References 251

6.2 Light-Harvesting Complexes in the Purple Photosynthetic Bacteria R.J. COGDELL (With 4 Figures) 1 Introduction 252 2 The B-890 Class of Light-Harvesting Complexes (LH I Complexes) . . 253 Contents XIII

3 The B-800-850 Complex Class of Light-Harvesting Complexes (LH II Complexes) 255 4 General Remarks on the Structure of Bacterial Light-Harvesting Complexes 257 References 258

6.3 Phycobilisomes E. GANTT (With 4 Figures) 1 Introduction 260 2 Stability of Phycobilisomes and Isolation 261 3 Structure and Composition 262 4 Reconstitution of Phycobilisomes 266 5 Association with Photosystem II 266 References 268

6.4 Light-Harvesting Pigment-Protein Complexes of Algae J.M. ANDERSON and J. BARRETT (With 6 Figures) 1 Introduction 269 2 Light Environment 269 3 Distribution of Light-Harvesting Pigments 269 4 The Structure and Spectra of Light-Harvesting Pigments 270 5 Pigment-Protein Complexes 272 6 Core Chi a-Proteins of PS I and PS II 273 7 Light-Harvesting Complexes of the Chlorophyta 274 7.1 Siphonales 275 7.2 Prochlorophyta 276 8 Light-Harvesting Complexes of the Chromophyta 276 8.1 Phaeophyta 277 8.2 Diatoms 279 8.3 Chrysophyceae and Xanthophyceae 279 8.4 Cryptophyta 280 8.5 Peridinin-Chl a-Proteins 280 8.6 Molecular Topology of Algal Pigment-Protein Complexes 281 8.7 Structure of Algal Thylakoid Membranes 281 9 Spatial Organization of PS II and PS I in Algal Thylakoid Membranes 282 10 Strategies of Light-Harvesting in Algal Groups 282 11 Conclusions 284 References 284

7 Trapping Events in Light-Harvesting Assemblies

7.1 Theory and Modeling of Excitation Derealization and Trapping R.S. KNOX 1 Introduction 286 1.1 Excitation Derealization and Trapping: Observations 286 1.2 Historical Development of Theory and Modeling 287 2 Derealization and Pairwise Transfer 288 2.1 States Produced by Light Absorption 288 2.2 Förster Theory of Pairwise Transfer 290 2.3 Application : Annihilation of Excitons 291 2.4 Beyond the Förster Theory : Coherence, Triplets, and the Monopole Approximation 292 3 Excitation Flow to Traps 293 XIV Contents

3.1 Principles : Adaptation of Pairwise Transfer Theory 293 3.2 Applications to Photosynthesis 295 4 Conclusions and Prospects 296 References 297

7.2 Excited State Kinetics of Chlorophyll Antenna Pigments A.R. HOLZWARTH (With 2 Figures) 1 Introduction 299 2 Instrumentation 299 3 Room Temperature Fluorescence Kinetics 301 4 Measurements of Thylakoid Fragments 305 5 Low Temperature Fluorescence Kinetics 306 6 Origin of the Long-Lived (Nanosecond) Decay 306 7 Conclusions and Outlook 307 References 308

7.3 Topology and Dimensions of Antenna Pigment Systems Probed by Exciton Annihilation N.E. GEACINTOV and J. BRETON (With 1 Figure) 1 Introduction 310 2 Principles of Annihilation Techniques 310 3 Exciton Annihilation and Effects of Domain Size 311 3.1 Small Domains 311 3.2 Large Domains 312 4 Annihilation and Capture by Reaction Centers: Topological Effects . . 313 5 Mobile Excitons Versus Localized Excitons 315 6 Conclusions 316 References 317

7.4 Molecular Orientation of the Pigments and the Problem of Energy Trapping in Photosynthesis J. BRETON (With 2 Figures) 1 Introduction 319 2 Orientation of the Absorption Dipoles in Vivo 320 3 Orientation of the Emission Dipoles in Vivo 322 4 Assignment and Molecular Origin of the Fluorescence Bands in Vivo 324 References 326

7.5 Excitation Transfer in Phycobiliproteins H. SCHEER (With 2 Figures) 1 Introduction 327 2 Structure of Phycobiliproteins 327 3 Photophysics and Photochemistry of the Chromophores 330 4 The Energy Transfer Chain 331 5 Dynamics of Energy Transfer 332 6 Physiological Status 334 References 336

8 Physical Determinants of Charge Separation Processes

8.1 Primary Electron Transfer Reactions in Photosynthetic Bacteria: Energetics and Kinetics of Transient States W.W. PARSON and D. HOLTEN References 342 Contents XV

8.2 Redox Center and Secondary Protein Structure Organization in

Bacterial Reaction Centers and the Cytochrome b/ct Complex D.M. TIEDE (With 2 Figures) 1 Introduction 344 2 Redox Components: Definitions, Abbreviations 344 2.1 Redox Centers in the Reaction Center 344 2.2 Redox Centers in the b/cj Complex 346 3 Reaction Center Structure 346 3.1 Protein Secondary Structure 346 3.2 Redox Center Locations in the Membrane Profile 346 3.3 Redox Center Separations 347 3.4 Redox Center Orientations with Respect to the Membrane Normal 348 3.5 Angles Between Donor and Acceptor Molecular Planes 349 4 Structure of the Cytochrome b/cj Complex 350 4.1 Location of Redox Centers in the Membrane Profile 350

4.2 Redox Center Separation: cyt bh—Q 350 4.3 Orientation of Redox Centers 351 References 351

8.3 Energy Trapping in Photosynthesis as Probed by the Magnetic Properties of Reaction Centers J.R. NORRIS and G. VAN BRAKEL (With 7 Figures) 1 Introduction 353 2 Mechanistic Aspects of the Photosynthetic Process 353 2.1 Key Features 353 2.2 Primary Photochemistry 353 2.3 Consequences of Blocking Secondary-Tertiary Chemistry 355 3 Components of the Reaction Center 356 3.1 Protein 356 3.2 Primary Donor 357 3.3 Acceptors 358 3.3.1 Bridging Intermediate and Primary Acceptor 358

3.3.2 Acceptor Complex QA — Fe — QB 358 4 Probing of Chemical Trapping by Magnetic Resonance Techniques . . 359 4.1 Introduction 359 4.2 ENDOR 360 4.3 EPR of Single Crystals 361 4.4 Optically Detected Magnetic Resonance (ODMR) 362 4.4.1 Introduction 362 4.4.2 ODMR of the Photosynthetic Triplet State, PR 363 4.4.3 Reaction Yield Detected Magnetic Resonance (RYDMR) of the Primary Radical Pair State, PF 364 5 Summary 367 References 367

8.4 X-Ray Diffraction Studies on a Crystalline Bacterial Photosynthetic Reaction Center: A Progress Report and Conclusions on the Structure of Photosystem II Reaction Centers H. MICHEL and J. DEISENHOFER (With 3 Figures) 1 Introduction 371 2 Results and Discussion 373 2.1 The Electron Density Map and Its Interpretation 373 2.1.1 General Description 373 XVI Contents

2.1.2 The Pigments and Their Arrangement 374 2.1.3 The Protein Part 376 2.2 Functional Aspects 377 2.3 Evolutionary Aspects 378 2.4 Implications on the Structure of PS II Reaction Center 379 References 380

8.5 Primary Structure and Function of the Reaction Center Polypeptides of Rhodopseudomonas capsulata - the Structural and Functional Analogies with the Photosystem II Polypeptides of Plants J.E. HEARST (With 2 Figures) References 388

8.6 Membrane Topology and Photochemistry of the Green Photosynthetic Bacterium Chloroflexus aurantiacus R.E. BLANKENSHIP and R.C. FULLER (With 4 Figures) 1 Introduction 390 2 The Chlorosome 391 3 Cytoplasmic Membrane 393 4 Reaction Center and Electron Transport Pathway 394 5 Reaction Center Pigment and Protein Composition 396 References 398

8.7 Optically Detected Magnetic Resonance (ODMR) of Triplet States in Vivo A. J. HOFF (With 7 Figures) 1 Introduction 400 2 Electron Spin Resonance of the Triplet State in Zero Magnetic Field . . 400 3 ODMR Techniques 402 4 Zero Field Splitting Parameters and Decay Rates 403 5 Relative Orientation of Optical Transition Moments and Spin Axes . . 404 6 Microwave-Induced Fluorescence (MIF) Spectra 407 7 Triplet-Minus-Singlet Absorbance Difference Spectra of Bacterial Reaction Centers 408 8 Linear Dichroic Triplet-Minus-Singlet Absorbance Difference Spectra of Bacterial Reaction Centers 413 9 Triplet-Minus-Singlet Spectra of the Green Plant Photosystems .... 418 10 Conclusions and Prospects 418 References 419

9 Photosystems I and II: Structure, Proteins, and Cofactors

9.1 The Reaction Center of Photosystem II B. A. DINER (With 1 Figure) 1 Introduction 422 2 Primary Reactants 422 2.1 Transmembrane Charge Separation 422 2.2 P-680 (also called Chi a„) 423 2.3 Pheophytin (Ph, also called I) 424

2.4 QA (also called Q, X-320, QH, Qi, A,) 424 2.5 Fluorescence 425 2.6 Redox Potentials 426 Contents XVII

3 Secondary Reactions, Donor Side 427 3.1 Reduction Kinetics of P-680 + 427 3.2 Signal II 428

3.3 Identity of Z (also called D1? Y) 428 3.4 Other Donors 430 3.4.1 Carotenoid 430 3.4.2 Cytochrome b-559 430 4 Alternate Acceptors 431

4.1 Low Potential Acceptor, QL 431

4.2 Q400 (also called AH, R and Q2) 432 4.3 a- and ^-Centers 433 4.4 Still More Heterogeneity 433 5 Polypeptides 433 References 434

9.2 Electron Transfer on the Oxidizing Side of Photosystem II: Components and Mechanism CF. YOCUM

1 Enzymatic Oxidation of H20 by PS II 437 2 Components of the Oxygen-Evolving Complex 438

3 Factors Influencing the Sequential Oxidation of H20 442 4 Summary and Conclusions 443 References 445

9.3 Proteins Participating in Photosynthetic Water Oxidation B. ANDERSSON (With 1 Figure) 1 Introduction 447 2 Extrinsic Proteins of Water Oxidation 447 2.1 The 33, 23, and 16 kDa Proteins. Identification and Connection with Water Oxidation 447 2.2 Properties, Biogenesis and Organization in the Thylakoids .... 450 2.3 Specific Functions in Water Oxidation 452 2.4 AIO kDa Polypeptide 453 3 Intrinsic Proteins of Water Oxidation 453 4 Hypothetical Architecture of the Water Oxidation Complex 454 References 455

9.4 Photosystem II Reaction Center: Polypeptide Subunits and Functional Cofactors C. J. ARNTZEN and H.B. PAKRASI (With 1 Figure) 1 Introduction 457 2 Polypeptides of the PS II Core Complex 457 2.1 51 kDa Polypeptide 459 2.2 45 kDa Polypeptide 459

2.3 QB (32 kDa) Protein 460

2.3.1 Quinone-Herbicide Interactions at the QB Site 461

2.3.2 Photoaffinity Tags of the QB Site 461

2.3.3 Mutations Affecting Quinone and Herbicide Binding at the QB Site 462 2.4 The 34 kDa D2 Protein 462 2.5 10 kDa Protein 463 3 Photosystem II : A Membrane-Protein Complex 464 References 466 XVIII Contents

9.5 The Mechanism of Photoinhibition in Higher Plants and Green Algae D.J. KYLE and I. OHAD (With 1 Figure) 1 Introduction 468 2 Phenomenology of Photoinhibition 468 2.1 Higher Plants 468 2.2 Algal Cells 469 2.3 Photoinhibition in Chloroplasts 470 3 Mechanism of Photoinhibition 470 4 Recovery from Photoinhibition Damage 471 5 Synergism Between Light and Other Stresses 472 6 Conclusion 474 References 474

9.6 Photosystem I Reaction Center and Its Primary Electron Transfer Reactions P. SÉTIF and P. MATHIS (With 2 Figures) 1 Introduction 476 2 The Primary Donor P-700 477 2.1 Basic Properties of P-700 477 2.2 Chemical Nature of P-700 477 2.2.1 Two Chlorophyll Molecules 479 2.2.2 One Chlorophyll Molecule 479 3 Photosystem I Electron Acceptors 480

3.1 Terminal Acceptors: FA and FB 481

3.2 Fx and A2 482

3.3 More Primary Electron Acceptors: A0, A, 482 4 Structural Aspects 483 4.1 Electron Carriers and Polypeptides 483 4.2 Geometrical Aspects 484 4.3 Functional Accessibility 484 5 Conclusion 484 References 485

9.7 Photosystem I Proteins F. -A. WOLLMAN 1 Introduction 487 2 The RC I Protein Complex 488 3 The LHC I Protein Complex 489 4 Architecture of the PS I Protein Complexes 492 5 Site of Synthesis and Assembly of Subunits of the PS I Protein Complexes 492 References 494

10 Energy Transfer and Energy-Coupling Processes

10.1 Composition and Structure of Cytochrome be, and b6f Complexes G. HAUSKA (With 2 Figures) 1 Introduction 496 2 Composition 497 2.1 Redox Components 497 2.1.1 Cytochrome b 497 2.1.2 Cytochrome C! or f 498 2.1.3 The Rieske FeS Center 498 2.1.4 Quinone 499 2.2 Polypeptides 500 Contents XIX

3 Structure 502 3.1 Genes and Primary Structures 503 3.2 A Topographical Model 504 References 506

10.2 Mobile Electron Carries in Thylakoids J. WHITMARSH (With 2 Figures) 1 Introduction 508 2 Diffusion in Two Dimensions - Definitions 509 3 Topology and Composition of the Thylakoid Membrane 513 4 Lateral Diffusion Between Appressed and Stroma-Exposed Membrane Regions 519 5 Lateral Diffusion Within a Localized Region 521 6 Concluding Remarks 524 References 525

10.3 Cytochrome b6/f Complex - Quinol Interactions P. JOLIOT and A. JOLIOT 1 Introduction 528 2 Slow Electrogenic Phase 528 3 Proton Translocation Associated with the Intersystem Electron Transfer 529 4 Reverse Electron Flow 530

5 Cytochrome b6/f Complex 530

6 Redox Potentials of the Carriers Involved in the Cytochrome b6/f Complex and in the Slow Electrogenic Phase 533 7 Models 534 References 538

10.4 Light-Driven Electron and Proton Transfer in the Cytochrome bei Complex R.C. PRINCE (With 2 Figures) 1 Introduction 539

2 Components of the Cytochrome bcx Complex 539 2.1 Cytochrome C! 539 2.2 Cytochromes b 540 2.3 The Rieske Iron-Sulfur Protein 540 2.4 Quinones 540

3 Stoichiometrics and Interactions in the bct Complex 541

4 Pathways of Electron Flow Within the bc{ Complex 542 5 Electrogenic Events in the be. Complex 544

6 Electron Flow in the bc{ Complex of Other Photosynthetic Bacteria 545

7 Light-Induced Electron Flow in the Mitochondrial bcL Complex . . . 545 References 546

10.5 Plastocyanin W. HAEHNEL (With 1 Figure) 1 Introduction 547 2 Synthesis 547 3 Structure 548 3.1 Refined Crystal Structure 548 3.2 Chemical Modification 550 4 Reduction of P-700 552 4.1 Surface Charges 552 XX Contents

4.2 Kinetics 552 4.3 Subunit III of PS I 553 5 Oxidation of Cytochrome f 554 6 Reactions in Situ 555 6.1 Organization of Electron Transport 555 6.2 Diffusion Coefficient 556 6.3 Kinetics in Situ 556 References 557

10.6 Local and Delocalized Interactions in Energy Coupling B.A. MELANDRI and G. VENTUROLI 1 Introduction 560 2 Steady-State Coupling in Chloroplasts 560 3 Steady-State Coupling in Bacterial Chromatophores 562 4 Transient Coupling Phenomena in Chloroplasts 563 5 Transient Coupling Phenomena in Chromatophores 564 6 ATP Synthesis Driven by Artificial Ion Gradients 565 7 Interpretation of the Phenomena of Localized Coupling 566 References 568

10.7 Proton-Membrane Interactions in Chloroplast Thylakoids R.A. DlLLEY 1 Introduction 570 2 Proton Uptake and Endogenous Buffering 570 3 Chemical Probes for Detecting Membrane Proton-Binding Groups . . 571 4 Sequestered Proton-Buffering Domains 572 5 Sequestered Proton Pools and Energy Coupling 574 References 575

10.8 Structure, Mechanism, and Regulation of the Chloroplast

+ H -ATPase (CF,-CF0) R.E. MCCARTY and CM. NALIN (With 1 Figure) 1 Introduction 576 2 Structure of CFI-CFQ 577 2.1 Subunit Stoichiometry 577 2.2 Organization of CF^CFQ 577 2.3 Roles of the Subunits 578 3 Mechanism and Regulation 579 3.1 Substrates 579 3.2 H+/ATP Stoichiometry 579 3.3 Regulation of Activity 579 3.4 Cooperativity During Catalysis 580

4 Genetic Analysis of CF,-CF0 581 5 Future Directions 581 References 582

10.9 Evaluation of Results on Nucleotide-Binding Sites of the ATPase Complex H. STROTMANN (With 1 Figure) 1 Introduction 584 2 Definitions 584 3 The Number of Nucleotide-Binding Sites 585 4 Localization of Nucleotide-Binding Sites 587 Contents XXI

5 Catalytic Mechanism 587 6 Roles of Tightly Bound Nucleotides 589 7 Conclusion 592 References 592

10.10 Sites of Herbicide Inhibition at the Photosynthetic Apparatus G. SANDMANN and P. BÖGER (With 2 Figures) 1 Introduction 595 2 Bioenergetic Reactions 596 2.1 Inhibition of Electron Transport Between the Two Photosystems 596 2.2 Inhibition of ATP Formation 597 3 Pigment Formation 597 3.1 Carotenoid Biosynthesis 597 3.2 Chlorophyll Biosynthesis 598 4 Peroxidation of Chloroplast Components 599 5 Conclusions 601 6 List of Herbicides Mentioned in the Text 601 References 602

11 Topography, Composition, and Assembly of Photosynthetic Membranes 11.1 Membrane Architecture of Anoxygenie Photosynthetic Bacteria S.G. SPRAGUE and A.R. VARGA (With 11 Figures) 1 Introduction 603 2 Green Bacteria 605 2.1 Morphology 605 2.2 Adaptation of the Photosynthetic Apparatus to Different Environmental Conditions 609 3 Heliobacterium chlorum 610 4 Purple Bacteria 610 4.1 Morphology 611 4.2 Development of ICM Structure 613 4.3 Additional Factors Influencing Membrane Structure 616 5 Summary 617 References 618

11.2 Topology of the Chromatophore Membranes of Purple Bacteria R. BACHOFEN and V. WIEMKEN (With 1 Figure) 1 Introduction 620 2 Topology of Pigment-Protein Complexes 621 3 Topology of Electron Carriers 626 3.1 Topology of the ATP Synthase, some Dehydrogenases and the Hydrogenase 627 4 Topology of Membrane Lipids 627 5 Conclusions 628 References 629

11.3 Synthesis and Assembly of Bacterial Photosynthetic Membranes T.J. DONOHUE and S. KAPLAN 1 Introduction 632 2 Assembly of the ICM in Steady-State Cells 633 XXII Contents

3 Induction of ICM Synthesis 634 4 Inhibition of ICM Synthesis 636 5 Repression/Derepression of ICM Synthesis 637 6 Future Prospects 638 References 639

11.4 Proteins of Cyanobacterial Thylakoids T.M. BRICKER, J.A. GUIKEMA, H.B. PAKRASI, and L.A. SHERMAN 1 Introduction 640 2 Protein Composition of Cyanobacterial Thylakoids 641 3 Chlorophyll-Protein Complexes 641 4 Photosystem II 644

5 Cytochrome b6/f Complex 649 6 Photosystem I 649

7 Fx and FrF0 650 8 Discussion 651 References 651

11.5 Surface Electrical Charges and Protein Phosphorylation J. BARBER (With 1 Figure) 1 Introduction 653 2 Surface Charge Densities 654 2.1 Particle Electrophoresis 654 2.2 Differential Doppler Micro Electrophoresis 655 2.3 Fluorescence Probe Techniques 655 2.4 Indirect Methods 656 2.5 Summary of Surface Charge Properties 656 3 Control of Photosynthetic Processes by Surface Charge 657 4 An Explanation of the Phosphorylation-Induced-Control of Energy Transfer in Terms of Electrostatics 660 5 Conclusion 662 References 663

11.6 Freeze-Fracture Studies of Mutant Barley Chloroplast Membranes D.J. SIMPSON (With 10 Figures) 1 Introduction 665 2 Location of Chlorophyll a/b Proteins 666 3 Location of the Photosystem I Reaction Center 669 4 Location of the Photosystem II Reaction Center 671 5 Reconstitution Studies 672 6 Effects of Single Gene Mutations on Functional Complexes 672 7 Concluding Remarks 673 References 674

11.7 Reconstitution of Thylakoid Components into Artificial Membranes I.J. RYRIE (With 1 Figure) 1 Introduction 675 2 Membrane Reconstitution - Overview, Techniques and Some Drawbacks 675 3 Reconstitutions with Thylakoid Proteins 676 3.1 Dicyclohexylcarbodiimide-Sensitive CF^FQ ATP Synthetase Complex 676 Contents XXIII

3.2 The Light-Harvesting Complex of Photosystem II, LHC II ... 677 3.3 Photosystem I 679

3.4 Cytochrome b6/f 679 3.5 Binary Protein Systems 680 4 Conclusions 681 References 681

11.8 Prolamellar Bodies C. LÜTZ (With 5 Figures) 1 Introduction 683 2 Formation of Prolamellar Bodies 684 3 Composition of Prolamellar Bodies 686 4 The Role of Lipids in Prolamellar Body Formation 688 5 Prolamellar Bodies and Membrane Formation — An Outlook .... 689 References 690

11.9 Acyl Lipids in Thylakoid Membranes: Distribution and Involvement in Photosynthetic Functions P.A. SlEGENTHALER and A. RA WYLER 1 Introduction 693 2 Aging of Thylakoids in Vitro 693 3 Extraction of Membrane Lipids by Organic Solvents 694 4 Modulation of Fatty Acid Composition . 694 5 Lipid Composition of Thylakoids and Subchloroplast Fractions ... 695 6 Modification of the Lipid : Protein Ratio in Thylakoid Membranes . . 697 7 Immunological Studies 699 8 Lipolytic Treatments 699 9 Biogenesis Studies 702 10 Conclusions 702 References 703

11.10 Biosynthesis of Chloroplast Membrane Lipids K. KEEGSTRA 1 Introduction 706 2 Structure of Chloroplast Membrane Lipids 706 3 Biosynthesis of Galactolipids 707 3.1 Synthesis of Prokaryotic Galactolipids 709 3.2 Synthesis of Eukaryotic Galactolipids 710 4 Biosynthesis of Other Chloroplast Lipids 711 5 Summary 712 References 712

11.11 Structural Properties and Molecular Organization of the Acyl Lipids of Photosynthetic Membranes D.J. MURPHY (With 5 Figures) 1 Introduction 713 2 Composition 713 3 Structures 714 3.1 Chemical Structures 714 3.2 Physical Structures 716 3.2.1 Diacylgalactosylglycerol 716 3.2.2 Diacyldigalactosylglycerol 718 3.2.3 Diacylsulfoquinovosylglycerol 718 XXIV Contents

3.2.4 Phospholipids 718 3.2.5 Thylakoid Lipid Mixtures 718 3.3 Physiological Significance of Lipid Structures 719 4 Asymmetric Distribution 719 4.1 Transverse Asymmetry 719 4.1.1 Antibody Studies 720 4.1.2 Lipolytic Studies 720 4.2 Lateral Heterogeneity 720 4.3 Extent and Functions of Lateral and Transverse Asymmetries . . 721 5 Lipid-Protein Interactions 722 5.1 Galactolipids 722 5.2 Phospholipids 722 References 725

Author Index 727

Subject Index 791 List of Contributors

JAN M. ANDERSON P. BÖGER CSIRO Lehrstuhl für Physiology Division of Plant Industry und Biochemie der Pflanzen P.O. Box 1600 Universität Konstanz Canberra City Postfach 5560 ACT 2601/Australia D-7750 Konstanz/FRG

B. ANDERSSON J. BRETON Department of Biochemistry Service de Biophysique University of Lund Département de Biologie P.O. Box 124 C.E.N. de Saclay S-22100 Lund/Sweden F-91191 Gif-sur-Yvette Cedex/France

T.M. BRICKER C. J. ARNTZEN Department of Chemistry E.I. du Pont de Nemours & Co. The University of Southern Mississippi Central Research and Development Hattiesburg, MS 39406-5043/USA Department Experimental Station R.J. COGDELL Bldg. 402, Room 3226 Department of Botany Wilmington, DE 19898/USA University of Glasgow Glasgow G12 8QQ/United Kingdom R. BACHOFEN Institut für Pflanzenbiologie J. DEISENHOFER der Universität Max-Planck-Institut für Biochemie Zollikerstraße 107 Abt. Strukturforschung II CH-8008 Zürich/Switzerland Am Klopferspitz D-8033 Martinsried/FRG J. BARBER Department of Pure and RA. DILLEY Applied Biology Department of Biological Sciences Imperial College Purdue University Prince Consort Road West Lafayette, IN 47907/USA London SW7 2BB/United Kingdom BA. DINER Institut de Biologie J. BARRETT Physico-Chimique Rockefeller University 13, rue Pierre et Marie Curie 1230 York Avenue F-75005 Paris, France New York, NY 10021/USA T.J. DONOHUE R.E. BLANKENSHIP Department of Bacteriology Department of Chemistry University of Wisconsin Arizona State University 1550 Linden Drive Tempe, AZ 85287/USA Madison, WI 53706/USA XXVI List of Contributors

P.L. DUTTON A.R. HOLZWARTH 607 Goddard Laboratories Max-Planck-Institut Department of Biochemistry für Strahlenchemie and Biophysics Stiftstraße 34-36 37th & Hamilton Walk D-4330 Mühlheim/Ruhr/FRG Philadelphia, PA 19104/USA

R.C. FULLER A. JOLIOT Department of Biochemistry Institut de Biologie University of Massachusetts Physico-Chimique Amherst, MA 01003/USA 13, rue Pierre et Marie Curie F-75005 Paris/France E. GANTT Smithsonian Environmental P. JOLIOT Research Center Institut de Biologie 12441 Parklawn Drive Physico-Chimique Rockville, MD 20852/USA 13, rue Pierre et Marie Curie N.E. GEACINTOV F-75005 Paris/France Chemistry Department New York University S. KAPLAN New York, NY 10003/USA Department of Microbiology University of Illinois J.A. GUIKEMA at Urbana-Champaign Division of Biology 407 South Goodwin Avenue Kansas State University Urbana, IL 61801/USA Ackert Hall Manhattan, KS 66506/USA K. KEEGSTRA W. HAEHNEL Department of Botany Westf. Wilhelms-Universität University of Wisconsin Münster Madison, WI 53706/USA Lehrstuhl für Biochemie der Pflanzen Hindenburgplatz 55 R.S. KNOX Department of Physics D-4400 Münster/FRG and Astronomy G. HAUSKA University of Rochester Institut für Botanik Rochester, NY 14627/USA Universität Regensburg Universitätsstr. 31 DJ. KYLE D-8400 Regensburg/FRG Martek Corporation 9115 Guilford Rd. J.E. HEARST Columbia, MD 21046/USA Department of Chemistry 430 Latimer Hall University of California C. LÜTZ Berkeley, CA 94720/USA Botanisches Institut der Universität Köln A.J. HOFF 1. Lehrstuhl Department of Biophysics Gyrhofstraße 15 Huygens Laboratory D-5000 Köln 41 of the State University NL-2300 RA Leiden/The Netherlands P. MATHIS D. HOLTEN Service de Biophysique Department of Chemistry Département de Biologie Box 1134 C.E.N. de Saclay Washington University F-91191 Gif-sur-Yvette Cedex/ St. Louis, MO 63130/USA France List of Contributors XXVII

R.E. MCCARTY W.W. PARSON Section of Biochemistry Department of Biochemistry SJ-70 Molecular and Cell Biology University of Washington Cornell University Seattle, WA 98195/USA Ithaca, NY 14853/USA R.C. PRINCE B.A. MELANDRI Exxon Research University of Bologna and Engineering Co. Institute of Botany Annandale, NJ 08801/USA Via Irnerio 42 1-40126 Bologna/Italy A. RA WYLER Laboratoire de H. MICHEL Physiologie végétale Max-Planck-Institut Université de Neuchâtel für Biochemie Ch. de Chantemerle 20 Abteilung Membranbiochemie CH-2000 Neuchâtel/Switzerland Am Klopferspitz D-8033 Martinsried/FRG IJ. RYRIE DJ. MURPHY Department of Applied Department of Botany Mathematics University of Durham Research School of Durham, DH1 3LE/United Kingdom Physical Sciences Australian National University CM. NALIN Canberra 2601/Australia Department of Cell Biology Roche Institute of G. SANDMANN Molecular Biology Lehrstuhl für Physiologie Roche Research Center und Biochemie der Pflanzen Nutley, NJ 07110/USA Universität Konstanz Postfach 5560 J.R. NORRIS D-7750 Konstanz/FRG Chemistry Division Argonne National Laboratory K. SAUER Argonne, IL 60439/USA Chemistry Department University of California I. OHAD Berkeley, CA 94720/USA Department of Biological Chemistry Institute of Life Sciences H. SCHEER The Hebrew University Botanisches Institut der Universität of Jerusalem Menzinger Straße 38 91904 Jerusalem, Israel D-8000 München 19/FRG D.R. ORT Department of Plant Biology P. SÉTIF 289 Morrill Hall Service de Biophysique University of Illinois Département de Biologie 505 S. Goodwin Ave. C.E.N. de Saclay Urbana, IL 61801/USA F-91191 Gif-sur-Yvette Cedex/ France H.B. PAKRASI Central Research and L.A. SHERMAN Development Department Division of E.I. du Pont de Biological Sciences Nemours & Co. University of Missouri-Columbia Wilmington, DE 19898/USA Columbia, MO 65211/USA XXVIII List of Contributors

P.A. SlEGENTHALER G. VAN BRAKEL Laboratoire de Physiologie Biokemi végétale Odense Universitet Université de Neuchâtel Campusvej 55 Ch. de Chantemerle 20 DK-5230 Odense M/Denmark CH-2000 Neuchâtel/Switzerland A.R. VARGA University of Illinois D.J. SIMPSON Department of Physiology at Urbana-Champaign Carlsberg Laboratory Department of Microbiology Gl. Carlsbergvej 10 Urbana, IL 61801/USA DK-2500 Copenhagen Valby/ G. VENTUROLI Denmark University of Bologna Institute of Botany S.G. SPRAGUE Via Irnerio 42 West Virginia University 1-40126 Bologna/Italy School of Medicine Department of Biochemistry J. WHITMARSH Morgantown, WV 26506/USA Department of Plant Biology 289 Morrill Hall L.A. STAEHELIN University of Illinois Department of Molecular 505 S. Goodwin Ave. Cellular and Developmental Urbana, IL 61801/USA Biology V. WlEMKEN University of Colorado Department of Botany Campus Box 347 University Boulder, CO 80309/USA Hebelstraße 1 CH-4056 Basel/Switzerland H. STROTMANN Botanisches Institut F.-A. WOLLMAN der Universität Düsseldorf Service de Photosynthese Universitätsstraße 1 Institut de Biologie D-4000 Düsseldorf/FRG Physico-Chimique 13, rue Pierre et Marie Curie J.P. THORNBER F-75005 Paris/France Department of Biology CF. YOCUM University of California Division of Biological Sciences 405 Hilgard Ave. The University of Michigan Los Angeles, CA 90024 Ann Arbor, MI 48109-1048/USA USA H. ZUBER D.M. TlEDE Institut für Molekularbiologie Chemistry Division und Biophysik Argonne National Laboratory ETH-Hönggerberg - HPM Argonne, IL 60439/USA CH-8093 Zürich/Switzerland 7.5 Excitation Transfer in Phycobiliproteins

H. SCHEER

1 Introduction

One of the major strategies in the competition of photosynthetic organisms for light is qualitative and quantitative adaptation in their light-harvesting appa• ratus (THORNBER, Chap. 3 ; STAEHELIN, Chap. 1 ; ANDERSON, Chap. 6.4, all this Vol.). This includes, in particular, the usage of pigments which harvest light efficiently in the spectral regions not covered by the chlorophylls a and b present in higher plants, e.g., the bacteriochlorophylls absorbing well beyond 700 nm (COGDELL, Chap. 6.2, this Vol.), and the phycobiliproteins and carotenoids absorbing in the "chlorophyll trough" between 470 and 630 nm (GANTT, Chap. 6.3, this Vol.). Among these, the phycobiliproteins are to date probably the best understood. They are not only the major light-harvesting pigments of cyan• obacteria, red algae, and cryptophytes (see STAEHELIN, Chap. 1, this Vol., for a general characterization of these species), microorganisms which are responsi• ble for a large fraction of the net photosynthetic production on earth, but they are also readily accessible and pleasing to the eye. Different aspects of phycobiliprotein research (GANTT 1979, 1981; GLAZER 1980, 1983; MACCOLL 1982; RÜDIGER 1979; SCHEER 1981, 1982; WEHRMEYER 1983; ZUBER 1978) and related topics (BRASLAVSKY et al. 1983; PRATT 1978, 1982; RÜDIGER 1980; RÜ• DIGER and SCHEER 1983) have been reviewed in the past 5 years. In this volume, GANTT (Chap. 6.3) describes the structure and function of phycobilisomes and Zu BER (Chap. 6.1) the primary structure of phycobiliproteins. This chapter pro• vides a brief overview of the energy transfer in biliproteins. Due to space limita• tions, only few citations could be incorporated. They are a biased selection from a large body of work, and should be used as a source for further references.

2 Structure of Phycobiliproteins

Most phycobiliproteins contain one or both of the chromophores, phycocyanobilin (Fig. 1 a) or phycoerythrobilin (Fig. 1 b), but additional chromophores of yet unknown structure are present, for example, in red algae, phycoerythrins ("phycourobilin"), in phycoerythrocyanins, and in particular in biliproteins from cryptophytes. Both chromophores are linked to their apoproteins via one, and in some cases two (RAPOPORT and GLAZER 1984) stable thioether bond(s) 328 H. SCHEER:

Fig. 1. Chromophores of phycobiliproteins. Molecular structures of a phycocyanobilin, the chromophore of PC and APC, b phycoerythrobilin, the main chromophore of phycoerythrins. A hypothetical conformation of the native phycocyanobilin in PC is shown in c (see SCHIRMER et al., 1985, for three dimensional structures of PC chromophores) at rings A or D to cystein residues, as has been inferred from several independent lines of evidence (RÜDIGER 1979; LAGARIAS et al. 1979; ZUBER 1978; KÖST-REYES and KÖST 1979; FREIDENREICH et al. 1978), including a recent X-ray structure (SCHIRMER et al. 1985). Additional, more labile bonds have been postulated to exist based on conflicting results (see SCHEER 1982). Studies with a new, mild chromophore cleavage method have recently shown the absence of a second bond in two phycocyanins (KUFER et al. 1983). The properties of the native biliprotein chromophores are profoundly in• fluenced by noncovalent interactions with the protein (see SCHEER 1982). Both the intense visible absorption and fluorescence of the phycobiliproteins, which are crucial for their functions, are caused by these interactions, and are absent in the denatured pigments and in free chromophores (see below). The phycobili• proteins thus present a good example of "molecular ecology", e.g., the optimiza• tion of cofactors to specific functional needs by interactions with the apoproteins (SCHEER 1982). All biliproteins are composed of two to three chromophore-bearing subunits. To date, the primary structures are known for more than ten different subunits, 7.5 Excitation Transfer in Phycobiliproteins 329 including the a- and /J-subunits of all biliproteins from the cyanobacterium, Mastigocladus laminosus e.g. allophycocyanin (APC), phyocyanin (PC) and phycoerythrocyanin (PEC) (FÜGLISTALLER et al. 1983), and the red (?) alga, Cyanidium caldarium (APC and PC) (OFFNER and TROXLER 1983; see also ZUBER, Chap. 6.1, this Vol.). In the pigments from cyanobacteria and red alga, the a-subunit (%17kDa) bearing 1-2 chromophores and a slightly larger ß- subunit carrying 1-4 chromophores, are always present in a 1:1 molar ratio. Additional chromophore-carrying y subunits of generally much larger size are known in some APC's and in red algal PE's (phycoerythrin) in (

3 Photophysics and Photochemistry of the Chromophores

Denatured biliproteins, in which the chromophores are uncoupled from the protein, as well as free bile pigments of similar structures, have flexible chromo• phores which are predominantly present in solution in a cyclic-helical conforma• tion (SCHEER 1982; BRASLAVSKY et al. 1983). These conformers have absorptions of moderate intensity in the visible range (eœ 17,000 M-1 cm-1 at neutral pH) (see BRANDLMEIER et al. 1981 ), low fluorescence yields (0 < 10 " 3 see BRASLAVSKY et al. 1983) with corresponding fast excitation decays (SCHNEIDER, unpublished) and low phosphorescence intensities (LAND 1979). The deexcitation is governed by internal conversion, which is promoted by one or more of the following mechanisms: proton transfer (FALK and NEUFINGERL 1979; FRIEDRICH et al. 1981), vibrations, or rotations around single bonds (KUFER et al. 1983), and by photoreactions, e.g., to the labile 10 E isomers (BRASLAVSKY et al. 1983). This deactivation is only hindered if the above mechanisms are blocked, e.g., by a rigid fixation (KUFER et al. 1983), distortion of the chromophores (FALK and THIRRING 1980), or (photo)selection of more rigid and extended conformers (BRASLAVSKY et al. 1983). In these cases, the lifetimes of the excited states are increased to allow high yields of fluorescence, or photochemical reactions to products like the 4 E- or 15 E isomers which are stable at ambient temperatures. In the biliproteins, such conformers are predominant. The chromophores are probably rigidly fixed in an extended and twisted conformation near the surface of the protein (SCHEER 1982). This rather general picture has been de• tailed recently by the current X-ray analysis of a PC (SCHIRMER et al. 1985). The energetically unfavorable conformations must be stabilized by noncovalent interactions with the polypeptide chains (SCHEER 1982). The absorption in the visible range is thus increased by a factor of ^5, and fluorescence by several orders of magnitude to & >0.5 (see GRABOWSKI and GANTT 1978 a). The phyco• biliproteins are photochemically stable at ambient temperatures. It is, however, of interest to note that the photoreactive, light-sensing plant pigment phytochrome has a chromophore structure and protein interaction that resembles PC, and that partial denaturation renders phycobiliproteins to a certain extent photoreactive by partial uncoupling from the protein (BJÖRN 1979; BRASLAVSKY et al. 1983). In spite of the large number of chromophores present in phycobiliproteins, there are generally only moderate interchromophore interactions. Strong cou• pling between chromophores has been established for the pigments in crypto• phytes (HOLZWARTH et al. 1983; JUNG et al. 1980; KOBAYASHI et al. 1979), and may also be present in some of the cyanobacterial and red algal ones, but many of the isolated phycobiliproteins do not show good evidence of such coupling as judged from their CD spectra. However, aggregation is often paral• leled by the rise of S-shaped CD signals which are indicative of strong coupling. Since the chromophores are probably located at or near the surface of the native biliproteins, this would mean that the nearest neighbors in situ may not be the chromophores on the same subunits or heterodimers, but rather on different ones [see STAEHELIN, Chap. 1, this Vol. for a discussion of phycobilisome packing in vivo and the recent x-ray results of SCHIRMER et al. (1985)]. 7.5 Excitation Transfer in Phycobiliproteins 331

4 The Energy Transfer Chain

The participation of phycobiliproteins in photosynthesis and, in particular, their association with photosystem II was originally recognized by action spectro• scopy (EMERSON 1958). Energy transfer from biliproteins to chlorophylls has also been demonstrated in solution (FRACKOWIAK et al. 1979; KRASNOVSKII and EROKHINA 1969). The discovery of PB'somes (GANTT and CONTI 1966) marks the beginning of an understanding of phycobiliprotein function in cyanobacteria and red alga on a molecular basis. PB'somes collect light efficiently in the wavelength range between 480 and 630 nm, but their fluorescence is emitted almost exclusively (>95%) by a small fraction of a minor constituent, e.g.,

APC (Amax, absorption «665 nm, 2max, emission «680 nm). If attached to the photosynthetic membrane, the energy is transferred further to the chlorophyll- containing reaction centers in the membrane, preferentially to PS II (GANTT 1981). In the original PB'some model derived from electron microscopic and bio• chemical studies of the red alga, Porphyridium cruentum, a phycobilisome some• what resembles an onion cut in half: a core, made up of APC, is surrounded by layers of PC and, further out, PE. Although this model has been considerably refined by applying improved techniques and working with organisms having less complex phycobilisomes (see above), the basic elements have remained un• changed : there is a morphological ordering from PE to PC to APC when going from the periphery to the center of the PB'some (see Fig. 2 in GANTT, Chap. 6.3, this Vol.). This sequence corresponds perfectly to the decrease in excitation energy among the three pigments, which can thus form an energy transfer chain in which the fluorescence of any preceding member overlaps reasonably well with the absorption of the next one (Fig. 2). This model was refined by fluorescence polarization experiments first undertaken by TEALE and DALE (1970), which suggested part of an energy transfer chain to be present already within each individual biliprotein. The absorptions of PC's and PE's can be resolved into components of slightly different energies. These are assigned to individual chro• mophores of often the same molecular structure, but with slightly different absorption energies due to different interactions with the protein. The ones absorbing at shorter wavelengths act as sensitizers (s) to the ones absorbing and fluorescing at longer wavelengths (f chromophores), which provides an efficient fine-tuning of the energy transfer. In cyanobacteria, each individual pigment represents only a rather short fragment of such an energy transfer chain. PC has, for example, one s chromo• phore on the /?- and one f chromophore on both the a- and /?-subunits. The full transfer chain is completed by the aggregation with APC and PE in the PB'some. The cryptophytes represent the other extreme. They do not have PB'somes, instead they have biliproteins which carry sufficiently different types of chromophores to establish an energy transfer over the full absorption spec• trum of biliproteins. The red algae, finally, use a combination of both. They have PB'somes in which the major biliproteins already span a wide absorption range. 332 H. SCHEER:

Chromophore Absorption Fluorescence Type * *inax \ax 3 500 — 515 2 (s) 545 ^ 560 2 (f) 570 — 585

1 (s) 590 — 605 1 (0 620 ^ 635

650 — 665

Chlorophyll aj 686 -h» Photooxidation CD

Fig. 2. Energy transfer scheme of a phycobilisome. Righi a diagramatic representation of a PB'some with only one outer rod containing B-PE (top double disk) and C-PC (lower double disk). Vertical arrows correspond to longitudinal energy transfer, with the downward direction strongly favored due to a larger overlap integral; horizontal arrows to transversal energy transfer within the disks. Left approximate absorption and fluores• cence maxima of the chromophores. *: 1 = phycocyanobilin, 2 = phycoerythrobilin, 3 = phycourobilin

The energy transfer chain is completed by the transfer to the chlorophylls within the photosynthetic membrane. Both the final donor in the PB'some and the acceptor in the membrane have not yet been firmly established. At present, the best candidates seem to be a large chromopeptide of the APC core (GINGRICH et al. 1983; REDLINGER and GANTT 1982; RUSCHKOWSKI and ZILINS- KAS 1982) and a chlorophyll protein close to or even identical with P-680, respectively (CLEMENT-METRAL and GANTT 1983, see also GANTT, Chap. 6.3, this Vol.).

5 Dynamics of Energy Transfer

A large fraction of the energy transfer in phycobiliproteins proceeds probably via a FÖRSTER type process (see SAUER, Chap. 2; KNOX, Chap. 7.1, both this Vol.). This has been deduced from the efficiency of the energy transfer on one hand, and the low occurrence of strong couplings on the other. GRABOWSKI and GANTT (1978 a, b) have critically investigated a series of biliproteins and calculated FÖRSTER'S critical distances, which are mostly in the range of 40-60 Â, 7.5 Excitation Transfer in Phycobiliproteins 333 well beyond the diameter of the phycobiliprotein subunits («30 Â). The formal• ism of FÖRSTER (1949) has been developed for a randomly ordered and fluctuat• ing system with high acceptor and low donor concentrations. Since probably none of these criteria applies to the phycobiliproteins (see above), a concise theoretical treatment of the process is not yet possible (BLUMEN and MANZ 1979; PEARLSTEIN 1982), but the critical radius is expected to be even larger based on still incomplete data on the chromophore orientations (see e.g., ZICKENDRAHT et al. 1980; SCHIRMER et al. 1985; GILLBRO et al. 1983). The sec• ond critical factor, i.e., the overlap integral between the donor fluorescence and the acceptor absorption, is also fine-tuned in the biliproteins (see above). Recent applications of picosecond time-resolved techniques to the energy transfer process in biliproteins have principally verified this concept. The first detailed investigations by SEARLE et al. (1978) established in Porphyridium omen• tum a sequential flow of excitation energy from PE via PC and APC to chloro• phyll. The observed rates of excitation energy transfer could be fit by the exp(t~1/2) decay law characteristic for the classical FÖRSTER tansfer. Although the actual kinetics can be fit well with a concise set of rise and decay times for the PB'somes of this organism (WENDLER et al. 1984), the physical laws governing the decay have been questioned. The better signal-to-noise ratios of the current experiments still do not allow an unequivocal discrimination between multi-exponential £Aexp(kt)] and non-exponential decay laws, but generally favor the former. This is in particular supported by the observation of corresponding decay and rise time constants in pairs of donors and acceptors, respectively (WENDLER et al. 1984). Qualitatively similar results have been obtained with entire PB'somes, as well as with isolated phycobiliproteins and their subunits from several cyanobac• teria and red algae (HEFFERLE et al. 1984a, b; HOLZWARTH et al. 1982; Ko- BAYASHi et al. 1979; PELLEGRINO et al. 1981 ; SUTER et al. 1984; SWITALSKI and SAUER 1984; WONG et al. 1981 ; YAMAZAKI et al. 1984). Photosynthetic antenna systems are optimized for high absorption cross- sections and cooperativity. They thus present a problem for the interpretation of time-resolved measurements, because *S— lS annihilation can compete effi• ciently with energy transfer (see BRETON and GEACINTOV 1980; GEACINTOV and BRETON, Chapter 7.4). This process has been demonstrated in phycobiliproteins and investigated in detail by ALFANO'S group (ALFANO 1982; PELLEGRINO et al. 1981; WONG et al. 1981). It can be distinguished from conventional excitation energy transfer (!S —°S) by its dependence on the intensity of the exciting light. More recent studies have thus focused on techniques which use only low photon fluxes, e.g., repetitive streak cameras (HEFFERLE et al. 1984a, b) or single-photon timing (HOLZWARTH et al. 1982; WENDLER et al. 1984; YAMAZAKI et al. 1984; HOLZWARTH, Chap. 7.3, this Vol.). An advantage of the former is the ready availability of polarized data, whereas the latter has an exceptionally high dy• namic range and can thus detect even minor contributions. Several criteria have been used to identify energy transfer in biliprotein antenna systems. The first is the analysis of time-resolved depolarization. Due to their rigid binding to proteins, rotational depolarization is generally thought to be absent on the subnanosecond time scale, and depolarization is thus be- 334 H. SCHEER:

lieved to be due to energy transfer among chromophores (GILLBRO et al. 1983; HEFFERLE et al. 1983, 1984a, b). It should be noted, however, that segmental movements of the chromophores (HEFFERLE et al. 1984 b) and nonparallel excita• tion and emission dipoles (SWITALSKI and SAUER 1984) have been implicated to rationalize the kinetics of some isolated biliproteins and their subunits. It is also not trivial to obtain energy transfer time from the raw data (HEFFERLE et al. 1984 a). The second criterion is the correspondence of decay time constants in the donors with rise time constants in the acceptors, which are separated by wavelength selective excitation and emission (HOLZWARTH et al. 1982; WENDLER et al. 1984; YAMAZAKI et al. 1984). The following conclusions can be drawn from the results obtained until now: the energy transfer from PE to the terminal APC chromophore proceeds at a time scale <100ps (e.g., 60-70 ps in Porphyridium cruentum, WENDLER et al. 1984; YAMAZAKI et al. 1984). It follows the pigment sequence expected from their energetic and morphological ordering within the PB'somes. The ener• gy transfer from PE to PC appears to be faster than that from PC to the APC core (SUTER et al. 1984). In isolated biliproteins, the transfer times increase with decreasing aggregation, e.g., from 70 ps in the trimer (ccß)3 to %500ps in monomeric (ay?)! PC from Mastigocladus laminosus (HEFFERLE et al. 1984b). The transfer times are also dependent on the species, the isolation procedure and temperature. An intriguing conclusion might be drawn from the comparison of isolated biliproteins with phycobilisomes, e.g., that the physiologically impor• tant longitudinal energy transfer toward the APC core is faster than the poten• tially wasteful transversal transfer within the platelets of the phycobilisome. Currently, kinetic data are only available for a single cryptophytan pigment, i.e., PC-645 from Chroomonas sp. (HOLZWARTH et al. 1983; KOBAYASHI et al. 1979). The findings support again a somewhat different behavior of the crypto• phytan pigments compared to the PB'some-forming biliproteins. Differences between the absorption and fluorescence excitation spectra have been discussed in terms of a heterogeneous population of f chromophores, albeit with the same decay kinetics, and the results support the presence of strongly coupled chromophores (JUNG et al. 1980).

6 Physiological Status

Photosynthetic organisms are dynamic systems with the capacity to react to environmental changes. These changes include the structure of the phycobil• isomes, and their light-harvesting and energy-transfer capacities. The best inves• tigated feature is the chromatic adaptation of many cyanobacteria, which in• volves changes in the composition and structure of the phycobilisomes in re• sponse to changes in the qualitity of light (BJÖRN 1979; GANTT 1981; GLAZER 1983). Other important factors are the nutritional and developmental status of the organisms (CSATORDAY 1978). Considerably less is known about the changes in the energy transfer kinetics in these systems. Using fluorescence spectroscopy CSATORDAY (1978) has followed the repigmentation of Anacystis 7.5 Excitation Transfer in Phycobiliproteins 335 nidulans cells after nitrogen starvation. This approach allows for the detection of particular biliproteins before they have been incorporated into the antenna system. HARNISCHFEGER and CODD (1978) have observed a decrease in biliprotein fluorescence after illumination of dark-adapted cells of the same organism, which was correlated with an increased chlorophyll fluorescence. Changes of the fluorescence yields (FORK et al. 1982) and kinetics (KARUKSTIS and SAUER 1984) upon treatment of algal cells with DCMU or desiccation, indicate the involvement of different chlorophyll containing antennas, and possibly also different PS II populations. This changed coupling between phycobilisomes and membrane-bound pigments probably contributes to the chromatic transients in cyanobacteria as well. These few examples have been chosen to illustrate that the physiological status of the cells (as well as the isolation procedures of PB'somes and their components) may produce significant changes in energy transfer reactions, which must be considered in the interpretation of the physico- chemical data.

Acknowledgements. The cited work of the author was supported by the Deutsche For• schungsgemeinschaft. I thank Drs. A.R. HOLZWARTH and S. SCHNEIDER for critically reading the manuscript.

Notes Added in Proof: The field has been advancing rapidly after finishing the manuscript for this mini review. Important aspects should be referenced. The crystal structure has been determined for a second phycocyanin, viz. from Agme- nellum quadruplicatum, which is hexameric instead of trimeric, but otherwise very similar to that of M. laminosus (T. SCHIRMER, W. BODE, M. SCHNEIDER, W. BODE, M. MILLER, M.L. HACKERT, J. Mol. Biol., in press). The details of chromophore structures and the linkage to the apoprotein have been determined by chemical methods, including the high-resolution NMR of bilipeptides (J.O. NAGY, J.E. BISHOP, A.V. KLOTZ, A.N. GLAZER, H. RAPOPORT, J. Biol. Chem. 260, 4864-4868 (1985)), milder degradation meth• ods (W. KUFER, G. SCHMIDT, O. SCHMID, H. SCHEER, Z. Naturforsch., in press) and sequencing studies (W. SIDLER, B. KUMPF, H. ZUBER, W. RÜDIGER, Abstr. Vth. Int. Symp. on Photosynthetic Procaryotes (ed. H. ZUBER, p. 303, 1985, ETH Zürich). The results include the structure of the phycourobilin chromophore in R-PE and the character• ization of singly and doubly bound chromophores, depending on the species and the chromophore attachment site. An assignment of spectrally distinct chromophores to the different chromophore sites of PC from M. laminosus has been suggested on the basis of fluorescence polarization, circular dichroism and photochemical reactivities (M. MIMURO, P. FÜGLISTALLER, R. RÜMBELI, H. ZUBER, Abstr. Vth. Int. Symp. on Photosynthetic Procaryotes (ed. H. ZUBER, 1985, p. 276, ETH Zürich; W. JOHN, R. FISCHER, S. SIEBZEHNRÜBL, H. SCHEER, in Anten• nas and Reaction Centers of Photosynthetic Bacteria - Structure, Interactions and Dy• namics (eds. M.E. MICHEL-BEYERLE, H. SCHEER, S. FISCHER, Springer, Berlin - Heidelberg - New York, in press). Several new results have been presented on the excited state dynamics in biliproteins. Fast (7-10 ps) decay times of the short-wavelength fluorescence and a corresponding rise-term of the longer wavelength fluorescence has been demonstrated for PC 612 from the cryptophyte, Hemiselmis virescens (CA. HANZLIK, L.E. HANCOCK, R.S. KNOX, D. GUARD-FRIAR, R. MACCOLL, J. Luminescence 34,99-106 (1985). All groups have adopted the multi-exponential decay analysis, at least as a phenomenological approach, requiring up to 5 components depending on the system and on the signal-to-noise ratios of the data. The physical correctness of this approach has been supported by a global analysis of data sets obtained at different excitation and emission wavelengths with a single set of components (A.R. HOLZWARTH, in Antennas and Reaction Centers of Photosynthetic Bacteria - Structure, Interactions and Dynamics (eds. M.E. MICHEL-BEYERLE, H. SCHEER, 336 H. SCHEER:

S. SCHNEIDER), Springer Verlag, Heidelberg, Berlin, New York, 1985, in press). This method allows at the same time the extraction of time-resolved spectral information. First data on the influence of linker peptides have shown only relatively small changes in the excited state dynamics of an R- and a C-PC (S. SWITALSKI, K. SAUER, pers. communication, J. WENDLER, W. JOHN, H. SCHEER, A.R. HOLZWARTH, unpublished re• sults). The spectra of the individual components contributing to the emission have been studied in a chromatically adapting cyanobacterium, Fremyella diplosiphon (M. MIMURO, I. YAMAZAKI, T. YAMAZAKI, Y. FUJITA, Photochem. Photobiol. 41, 597-603 (1985). Partic• ular attention has been paid to the relative importance of lateral and vertical energy transfer, with the latter being favored in green-light adapted cells containing PE, PC and APC. The uncoupling of phycobilisomes from the PSII-related chlorophylls has been induced by heat stress in Anacystis nidulans (P. MOHANTY, S. HOSHINA, D.C. FORK, Photochem. Photobiol. 41, 589-596 (1985). In an interesting application, tandem phycobiliprotein conjugates have been used as immunofluorescent probes with an unusually large Stoke's shift due to the energy transfer from PE to APC (A.N. GLAZER, J. STRYER, Biophys. J. 43, 383-386 (1983)).

References

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Subject Index

A(FA) 481 chlorophyll a 325

A2 482 complex, bacterial 252, 257 absorbance spectra of bacteria 99 isolation 245 of bacterial reaction centers 120 effect 287 of siphonaxanthin-Chl a/b-proteins 275 LH I 252, 257 absorption, band broadening 90 LH II 252, 257 cross-section 88 mobile 48 determining factors 319 photosystem I 152, 170 magnetic resonance 401 photosystem II 264 acclimation, low temperature 37 pigments 299 Acetabularia 2 anthracene-tetracene model system 287

acyl lipids 693-703 antibodies, cytochrome b6/f 42

chlorophyll-protein complexes 696-705, 713 cytochrome C2 626 lateral heterogeneity 697, 703 H, L, M subunits 622 thylakoid membrane 693-705, 713 lipid 699, 720 transverse distribution 699, 706 Nostoc 265 unsaturation 695 oxygen-evolving system 451 adenylate sulfate 217 antioxidant 470 ADMR 363, 401, 409, 410 appressed membrane regions 720 ADP sulfurase 217 asymmetry, lipid 719 ADRY reagent 430 ATP synthase 42-44, 229-230, 650 aging of thylakoids 393 activation 584 algae, chloroplast membrane organization 6, binding 588, 590 53-61 coupling 229 Chromophyta 58-61 inactive-active transition 567 eukaryotic, systematic organization 599 phosphoryl transfer 588 evolution 8 stereochemistry 588 green 299 stoichiometry 229 pigment-protein complexes 116 structure 229 red 26,53,241,327 topology 627 thylakoid membranes 281 ATP synthesis 564, 576 allophycocyanin 241 artificial ion gradients 565 Alocasia 24 flashing light 564 alternating site hypothesis 187 inhibition 597 Amaranthus 25, 597 lag 574 amino acid sequence 132, 231, 243 ATPase (see ATP synthase) 9-aminoacridine 655 ATP-binding (see nucleotide binding) amphipathic protein 150 atrazine 24, 597 Anabaena 53, 173, 263, 496 A triplex 25 Anacystis 261, 335, 649 azidoatrazine 379, 461 annihilation, of excitons 291 antenna (see also light harvesting complexes) 89, 98-135 B(Bulk)-820, nomenclature 252 ^-apoproteins 258 B-790 392 ^-carotene 13 B-790, chlorosome 392 792 Subject Index

B-800-820, antennae 125, 245, 257 biliproteins, aggregation 329 protein 107, 124 binding site, chromophore 244, 249 spectral analysis 99 bipyridyliums 595 B-800-850, antennae 107, 123, 125, 245 blue light, prolamellar body 690 nomenclature 252 bridging intermediates 355 protein structure 133 brown algae 58-61,270-273,276-282 spectral analysis 99 buried domain, lumen pH 572 B-800-850 protein comparison 124

B-850-protein 123 C4 pathway 4

B-875-890, antennae 246 C4 thylakoid morphology 25 B-875-protein 123, 125 calcium 438, 452, 453 B-890-spectral analysis 99 calmodulin antagonists of photosystem II 649 B-890-protein 107, 123, 124, 125, 132 carbanilates 595 structure 133 carotenes, green bacteria 126 carotenoid-Chl a/c-protein 277 b-cycle 176 carotenoids 101, 106 Bacillariophyceae 6 algae 116 bacteria, green 197-232, 371, 390-399, 605-610 B-890-protein 99 chlorophyll-complexes 126 bacteria 119, 122, 253, 398 bacteria, purple, alpha-apoproteins 258 biosynthesis 597-598 photosynthetic membranes 197-232, 604, etioplasts 586-598 610-617, 620-630, 632-638 mutants 625 reaction centers 113, 119-125, 457, 458 catalytic hydrogénation, fatty acyl chains 694 bacteriochlorophyll a, antennae 246 catalytic mechanism 587-589 bacteriochlorophyll a-protein, green bacteria Caulerpa 10 102, 126, 132, 133 CC I (see also chlorophyll protein complexes) bacteriochlorophyll a, purple bacteria 119-125 105, 108-111, 116, 118, 119, 130, 135 bacteriochlorophyll anion 354 CC II (see also chlorophyll protein complexes) bacteriochlorophyll b, absorbance 102 105, 108, 112, 113, 116, 118, 130

bacteriochlorophyll, B-890 253 CF{ (see coupling factor and ATP synthase)

fluorescence 624 CF0 (see coupling factor and ATP synthase) photon absorbtion 338 charge recombination, fluorescence 306 reaction center 93 Klimov mechanism 302 special pair 94 model 325 bacteriochlorophyll binding 382 pheophytin 307 bacteriochlorophyll-binding proteins 633, 638 charge separation 93, 94 bacteriochlorophyll c 126, 392 annihilation 354 bacteriochlorophyll cation 354 radical pair 353 bacteriochlorophyll d 126 charge transfer, reaction center 412 bacteriochlorophyll e 126 transitions 340 bacteriopheophytin, green bacteria 119, 121 zero field splitting 403 purple bacteria 119,121 chemiosomotic theory 179,180,181 reaction center 338 chilling injury 67, 718 barley 665 Chi a-P2 130 be, complex 208, 209 Chla-P3 130 BChl (see bacteriochlorophyll) bilayer, fluidity chi a/b LHC (see also light harvesting com• 155 plex) 31,35,44-47 integral membrane proteins 150, 152 chla/b-Pl 130 lipids 149, 157, 716 chi a/c 277, 278 bentazon 23 chi d 271

/?-carotene 269, 273 chlc2 271,280 antennae 272 Chlamydomonas 8, 13, 161, 471, 490, 668 orientation 484 chloramphenicol 472 photosystem I 109, 110, 273 Chlorella 2, 304, 305, 313

photosystem II 273 chloride, 02 evolution 438, 452 biliprotein organization 329 Chlor obiaceae, reaction center 208, 390-398, biliprotein, transfer time 334 604-610 Subject Index 793

Chlorobium 127, 390-398, 496, 604-610, 545 counter ions 653 chlorobium vesicles (see chlorosomes) coupling factor (see also ATP synthase) 42, Chloroßexacae 119, 121, 122, 126, 127, 150, 152, 160, 187 390-398, 604-610 chlorophyll ratio 580, 581 Chlorophyceae 6 cooperativity 580, 581 chlorophyll 104 distribution 42, 50 reaction center 93 genetic analysis 581 chlorophyll a 100 light activation 186 in algae 269, 273 lipid requirement 152 in reaction center I 110, 273 localization 158, 181 in reaction center II 273-274 molecular weight 586 orientation 319 nucleotide interactions 584 P-700 477 of photosynthetic bacteria 185 special pair 374 organization 577 surrounding chlorophyll b 114 per chlorophyll 152, 153, 182 chlorophyll a/b ratio 101, 114 PFu particles 43 chlorophyll b 100, 276 proton translocation 183, 576 linear dichroism 321 reconstituted 43, 676 chlorophyll biosynthesis 598, 702 regulation 579 chlorophyll c 58, 116 roles of subunits 578 chlorophyll Cj 271 structure 576, 577

chlorophyll c2 271 substrates 579 chlorophyll proteins (see reaction center and subunit stoichiometry 577 light-harveating complexes) CP (see also chlorophyll proteins) Chlorophyta 270, 274, 281, 282, 284 CP1 109 chloroplast, DNA 11 CP 29 44, 108, 113, 130 envelope 2-10, 706 CP 43 108, 112, 433, 459 evolution (see evolution) amino acid sequence 460 membrane components 35 CP 47 108, 112, 433, 459 morphology 1, 665 amino acid sequence 459 stroma 10-14 CP I 40, 103, 130, 483, 488, 493, 642 survival after isolation 10 CPI* 110 chlorosomes 126, 390, 391-393, 605, 606, 609 CP la 110 assembly 610 CPU 44, 103, 108, 130, 642 baseplate 607 CPU* 44 core 618 CP III 104, 108, 130, 433, 642 envelope 607 CP IV 104, 108, 112, 130, 433, 642 rod elements 607 CPV 642 chromatic adaptation 18, 264, 334 CPa 108, 130 chromatic transients 335 CPO 40, 112 Chromatium 122, 124, 256, 545 Cr3+ 551 chromatophores 88, 201, 245, 328, 562-565 cross-linkers, chemical 622 chromophyta 58-61, 270, 276-282 Cryptomonads 277 Chroomonas 334 Cryptomonas 59 Chrysophyceae 6, 279-280 Cryptophyceae 6, 280, 282, 327 circular dichroism, biliproteins 330 crystals of reaction centers 372, 625 B-890 254 cyanelles 28 P-700 479 Cyanidium 53, 57, 329

C02-fixation 468 cyanobacteria 15, 53, 116, 143, 270 Codium 10, 275 genes 640 coherence 292 lipid composition 150 conformational changes of protein complexes pigment-protein complexes 117-119 571, 590 photosystem I 169, 170 conformers 330 photosystem II 459 cooperativity in ATP synthase 586 phycobilisomes 241, 327 copper binding 548 structure 15, 17, 281, 282, 284 core complex I (see CCI) thermophilic 69 794 Subject Index

Cyanophora 26, 53, 268 diatoms 277, 279 cyclic-helical conformation 330 diffuse layer 653 cytochrome b 540 diffusion coefficients 177, 178, 510, 512, 513 cytochrome b-559 159, 160, 383, 444, 453, 458 constant of exciton 294 cytochrome b gene 503 Fick's laws 510

cytochrome bct complex 496, 497 Stokes-Einstein equation 510, 511 components 539 difunon 595 electron flow 539-546 digalactosyldiacylglycerol 696, 697, 699, 700, mitochondria 496, 545 702, 713 stoichiometrics 541-542 in prolamellar bodies 72, 687, 688 topology 626 in thylakoids 150-158, 178

cytochrome b6/f complex 40-42, 50, 145, 152, structure 151,713-725 160, 177, 178, 181, 496, 516-524, 528, 640, dimer of P-700 478 649 dinoflagellates 280 electron transport 172 Dinophyceae 6 freeze-fracture 174 diphenyl ethers 595 hydropathy plots 189 diphenylamines 595 localization 158, 532 dipole-dipole interaction tensor 400, 402 proteins 152 diuron 164 proton translocator 679 DNP-INT 595 Q-cycle 175, 176 Doppler shift 655 reconstitution 42, 679 double resonant 403

cytochrome b6/f complex ratio with photosys- DTP 595 tems 182 DTT 590 cytochrome c 220-228, 373, 378 heme structure 224, 225 edge particles 609 high-low potential forms 220, 227 EF particles 30-72, 665-673 cytochrome c-552 228, 394, 548, 650 EFs particles 32, 44, 668

cytochrome c{ 539-540 EFu particles 32, 44, 668 gene 503 electric field 85, 95

cytochrome c2 626 electrochemical potential 183 cytochrome f 160, 173, 174, 531, 533, electrochromic band shift 176 554-555, 649 electron acceptors 93, 338, 339, 480-483 cytochrome f topology 627 electron donors 93, 165-174 cytochrome f reaction with plastocyanin 555 electron flow, reverse 211 cytochrome oxidase 444 electron microscopy, freeze-fracture 1-72, 665-674, 718

D-l protein (see also QB protein) 385, 460 electron spin resonance 400-402 D-2 protein 372, 379, 385, 460, 462 electron transfer 161-162, 169-170, 200 DBMIB 174, 177, 595 electron transport 87, 92-93, 161-167, DCCD-binding protein 627 172-179 DCMU 596 electrical responses 347 decay laws 333 in bacteria 198-200, 338-342 decay rates 403, 405 in cytochrome be complexes 496 decay time constants 334 in photosystem I 476-485 delayed fluorescence 340 in photosystem II 447 derealization 286, 288, 292 inhibition 596-597 depolarization 334 long range 555 Deriphat 160, 105, 130 electron tunneling 344, 349, 350. desaturase complex 598 electrophoresis 98-135, 641 development 702 emission 303, 319 DGDG (see digalactosyldiacylglycerol) ENDOR 360-361,479 diacyl lipids 149, 677 endosymbiotic 5-10, 19, 143 diffusion 177 energetics 338 diacylglycerol synthesis 708, 709 energy conversion 87 diacylsulfoquinovosylglycerol 150, 151, 157, energy distribution, PS I and PS II 64-66, 657, 686, 696-699, 702, 713 660 Subject Index 795 energy transfer 238, 250, 331 surface charge 656 dynamics 332 yield 302

1S-1S annihilation 333 F0 (see coupling factor and ATP synthase) energy trapping 319-325 Forster transfer 92, 290, 332, 333 energy-linked binding change mechanism 588 FRAP 511 envelope membranes 2-10,708-711 freeze-fracture 1-72 EPR 224,438,479 nomenclature 30 ethylenediamine 551 of bacterial membranes 605-617 etioplasts 70-72, 683-691 of envelope membranes 9, 10 Euglenophyceae 6 of prolammellar bodies 71, 72 evolution 8, 197, 238-251, 265, 329, 371-380, of thylakoid membrane 20, 21, 30-72, 382-388 665-674 exchange energy 400 Fremyella 266 excitation derealization 286-297 fucoxanthin 116, 271, 272 excitation energy transfer 47, 48, 86, 90, 267, fucoxanthin-Chl a-proteins 116, 277-279 668 Fucus 60 excited state 88 Fx 482 exciton 90, 294, 412, 413 annihilation 310, 315 g=4.1 EPR signal 438 coupling 250, 410 galactolipid synthesis (see also mono- and diga• delocalized 90 lactosyldiacylglycerol) 707-711 lifetimes 313 galactose 715 splitting 289 galactosyl transferase 708 theory 479 gel phase 716 trapping 325 Glaucocystis 28 eyespots 13-14 Gloeobacter 19, 55, 263 glycolipid (see mono- and digalactosyldiacylgly• f chromophores 331,334 cerol) F-685 322 glycoproteins 154 F-695 322 Gonyaulax 61

Ft (see coupling factor and ATP synthase) Gouy-Chapman theory 653 Fi subunits 651 grana, formation 47 fast component 302 grana membranes 38, 114, 147-149, 155-157, fatty acids 24, 693, 700, 708, 710, 713 177, 653-663, 665 FDMIR 407 greening 493, 684, 702 FDMR 401, 410 Griffithsia 26, 261 Fe(II) binding 382 group transfer reaction 184, 187 Fe-S center 536 Fe-S protein 533, 537 H+/ATP stoichiometry 579 feedback mechanism, phosphorylation 66 H+/P ratio 561 ferredoxin NADP oxidoreductase 160, 168, H-ATPase (see also proton) 576 171, 172, 176, 177, 476, 650 H-subunit 121, 356, 376, 382-388, 397, 615, flavodoxin 650 629 fluidity of thylakoid membrane 67, 662, 694, H20 as a reduced substrate 213

695, 697, 698 H2S as a reduced substrate 213 fluometuron 23 heat stress 66-67 fluorescence 299,311,313,340 Heliobacterium 605, 610 decay 301, 313-316 Helmholtz layer 654 detected magnetic resonance 401 Hemiselmis 29 of chlorophyll 299,301,641 herbicide list 601 of photosystem I 305, 425-426 herbicide resistance 165, 695 of photosystem II 425-426 herbicides 23, 164, 165, 379, 461, 686 of phycobilisomes 331 electron transport 24, 462 photosynthetic capacity 468 sites of inhibition 465, 595-602 polarization 289, 331 heterocyst 53 quenchers 286, 315 hexagonal II phase 689, 713-724 spectra 320 19' hexanoylfucoxanthin 279 796 Subject Index

HgCl2 550 LHC (see also light harvesting complex) histidine 623, 625 LHC, chi a/b-protein 31, 35, 44^47, 108 homology in reaction center 385 LHC I 40, 50, 108-112, 116, 130, 274, 276 hydrogen bonding chains 571 absorption spectra 490 hydropathy indexes 660 electrophoretic mobility 497 hydrophobic markers 622 nomenclature 105 hydrophobic proteins 384 subunits 490, 497 hydrophobicity plots 134 LHC II 108, 113-116, 130, 133, 135, 21A-216 hyperfine coupling 479 excitation transfer 680 fluorescence 303, 304 ICM (intracytoplasmic membrane), aerobic nomenclature 105 growth 637 reconstitution studies 677 formation 613-615, 617, 636 structure 134 function and structure 634 LHCP (see also light harvesting complexes) induction 615, 635 108 LHCP1 130 02 and light control 632, 635 protein insertion 634 LHCP 2 130 synthesis 637 LHCP 3 130 Ilf 440 LHP 621 Ilvf 440 lifetimes 302 light activation 186 immunological techniques (see antibodies) induction, anaerobic 613, 615 light-harvesting apparatus, adaptation 327 membrane 618 light-harvesting complex 98-135, 641-644 inner bulk phases 574 bound 35 interchromophore interactions 330 chi a 325 intermittent light 303 chla/b 19,44,320 internal conversion 330 chi c-protein 277 intersystem crossing 402 development 100 intracytoplasmic membrane (see ICM) freeze-fractured 46 inverted micelles 689, 722 intermittent light 135, 303 iodination 641 isolation 128-131 IPI 474 lipid association 696-702, 724 membrane adhesion factor 45 iron-sulfur centers, PS I 171, 180, 181, 481 mobile 31, 35, 44, 50, 64, 65 iron-sulfur cluster, cyt b6/f complex 673 of algae 269-684 iron-sulfur proteins 643 isoelectronic 653 of bacteria 117,124, 238-250, 252-258, 618, 621 particles 66 KCN 550 PFs particles 668 kinetics of electron transfer reactions 338 polypeptides 161 kinetics, single turnover 221 protein 105 Klimov mechanism 301, 307 reconstituted 45 three-dimensional map 47 L-subunit 376 topology 621-626 lake model 312 light-harvesting, long term modulation lasers 310 283 lateral asymmetry 157, 158 light-harvesting pigment proteins 238, 246, lateral heterogeneity 35, 49, 177, 282, 720-721 250 lateral migration of membrane components light-harvesting pigments, accessory 14 61-64, 660, 661 light-harvesting polypeptides, alpha and beta lateral separation of membrane proteins 182, 250 662, 673 evolution 240 lauryl dimethylamine oxide 106 light-harvesting strategies 282 LD 319-321,424 light-harvesting systems, accessory 16 LDS-Page 641 linear dichroic triplet-minus-singlet absorption LH I 616,633 413-417 LH II 636 linear dichroism 319-321, 424 Subject Index 797 linolenic acid 24, 662, 698, 701 membranes, appressed 63, 656, 697 lipid, boundary 152 cytoplasmic 390, 393 lipid enrichment of thylakoid membranes 698 electrical properties 202 lipid extraction 694 envelope 2-10, 706, 710 lipid pools 703 extra-chloroplastic 5 lipid-protein interactions 693-705, 722 fluid mosaic model 620 lipid to protein ratio 697-699 inner envelopes 4 lipid transfer 710 intracytoplasmic 390 lipids (see also acyl lipids and mono- and diga- lateral heterogeneity 35, 49, 177, 282, 720, lactosyldiacylglycerols), eukaryotic 707 721 prokaryotic 707 non-appressed 14-25, 656, 697 surface charge 656 menaquinone 372, 375, 394 lipolytic treatment of thylakoid membranes metastable buffered proton pool 572 699-702 metastable proton pool 572 lipophilic anions 443 methabenthiazuron 23 liposomes 42, 38, 672 MGDG (see monogalactosyldiacylglycerol) liquid-crystalline phase 716 Micrasterias 2 lithium dodecyl sulfate 104 microsecond laser pulse 310 localized domains 570 microwave induced fluorescence spectra Lolium 20 407-408 low oxygen tension 613 microwave selection 413 L-subunit 121, 356, 376, 383-388, 397, 626, MIF 407,408 629 migration of exitation 286 luminescence 425 mitochondrial bcx complex 496 lutein 272 mitochondrial electron transfer 200 lutein-chl a/b proteins 274 mitochondrial evolution and symbiosis 197 lysine-specific protease 465 MK-616 595 modification, chemical 571 M+ 438 molecular ecology 328 m-phehoxybenzamides 595 molecular excitation, localized 90 M-subunit 121, 356, 376, 383-388, 397, 626, molecular triplet PR 367 629 monogalactolipid (see monogalactosyldiacylgly• macromolecules, assembly 638 cerol) magnesium ions, proton gradient 229 monogalactosyldiacylglycerol, biogenesis 702 magnetic fields 340, 341 degree of unsaturation 696 magnetic resonance technique 356, 400 in prolamellar bodies 72, 683-692 magnetophotoselection 402, 405 in thylakoid membranes 150-158, 696-700 manganese 438, 450, 668 structure 713-725 Master Equation 294 monolayer 721 Mastigocladus 57, 242, 329, 334 monopole approximation 292, 293 melittin 699 morphological organization of phycobilisomes membrane, adhesion factor 45 331 architecture 603 morphology of bacterial reaction center 420 asymmetry, lipid 719 Mossbauer spectroscopy 481 attachment site, chlorosomes 608, 609 multigene family of LHC II 114 fluidity 155 multiline signal 439 fracture faces, nomenclature 30 mutants, chlorophyll b-less 37, 45, 103, kinase 659 666-669 labeling 621 carotenoid 481 lipids, phase transitions 67, 155 herbicide resistance 462, 595-597 morphology 69 PS I 39,669 phosphatase 659 PS II 34, 35, 668 position of redox center 350 ultrastructure 665-674, 689 protein, amphipathic 150 water oxidation 450 reconstitution 34-50, 675 stacking 45, 63, 146, 148, 149, 154, 155, NAD+ reduction 206, 210 656, 697 NADP+ reduction 206, 212 798 Subject Index

nearest neighbor analysis of water-splitting en• pairwise transfer 288-293 zymes 451 particle electrophoresis 654, 655 Neoghardiella 267 particle size histograms 33, 666 Nerium 67 particles, intramembrane 34, 62, 665-674 neutron diffraction 624 PC (phycocyanin) 330

NH2 groups 574 PChl(ide) 686

NH2OH 437 PCR 689

NH3 437 perforated lamellae 689 nitrofen 595 peridinin 271, 272 non-heme iron 375 peridinin-Chl a-protein 117, 280, 281 nonappressed regions 720 peripheral reticulum 4 norflurazon 595 permeability of thylakoid membranes 156 Nostoc 263 peroxidation 599-601 nucleotide-binding sites 585-587 herbicide induced 600 nucleotides, exchangeable 586 pF 339 tightly bound 586, 591 radical pair 354 PF particles 30-72, 665-673 O-phenanthroline 376 pH in lumen 570, 572 Ochromonas 1 pH-clamp 572 octyl glucoside 129, 591 pH-gradient 186, 560 ODMR 362-367,400 pH-indicating dye 573 ohmic components 567 Phaeophyceae 6 one-electron intermediate 382 Phaeophyta 277 optical absorption cross-section 314, 316 phase transitions (see also lipids) 67-70, 155 optical transition dipole moments 348, 349 phehylhydrozones 595 orientation of optical transition moments pheophytin 93, 159, 162, 424-427 404-406 phonon wing 412 of pigment molecules 319-325, 484 Phormidium 118 overlap integral of biliproteins 333 phosphate group transfer 180, 187 oxadiazon 595 phosphate-water-oxygen exchange 589 oxidation rate 222 phosphatidylcholine 150, 151, 155, 696-703, oxidizing potentials of reaction centers 214 713 oxygen-evolving complex, 10 kDa protein 453 phosphatidylethanolamine 714 16 kDa protein 447 phosphatidylglycerol 150, 151, 157, 694-702, 711

23 kDa protein 447 phospholipase A2 701 32 kDa protein 447 phospholipase C 701 oxygen-evolving systems 231, 437-445, phospholipase D 701 447-456 phospholipid (see also lipids) 628, 686, 688, oxygen radicals 470 716, 677 oxygen regulation 635 insertion into ICM 616, 634 phosphorescence 401 P-430 171,481 phosphoryl group transfer 144, 145 P-680 112, 160, 162, 165, 170, 321 phosphorylating units 181, 182 dimer vs. monomer 423 phosphorylation, external electric field 565 P-680+ reduction kinetics 427 fluorescence 304 P-700 145,160,169-172,321,476-485, in Porphyridium 267 487-494, 669 kinetic analysis 565 of cyanobacteria 118,647,649,650 maximal affinity 562 P-700+ reduction kinetics 172, 175, 553, onset lag 574 556-557 rate 567 P-700, apoprotein 127 phosphorylation/dephosphorylation of pro• chlorophyll a-protein 103, 108, 109 teins 64, 115, 659 reduction by plastoxyanin 552-554 photoaffinity herbicides 461 P-870 121, 122 photobleaching 470 primary donor complex 357 photochemical reaction centers 106 P-960 121, 122 photoinhibition 461, 468-474 P/2e ratio 561 photon absorption 85 Subject Index 799 photon correlation length 289 distance to nearest neighbor 516, 518 photophosphorylation 179-189, 560, 561, 651 EF particles 34-38, 48, 671-672 photorespiration 469 electrogenic reaction 423 photoselection 349, 413 fluorescence 426 photosynthetic bacteria 15, 98, 102, 106, 107, fluorescence quenching 313 197-232, 252-258, 338-342, 344-351, freeze-fracture particles 32-63, 48, 55 371-380, 382-388, 390-398, 603-619, gene sequences 458 620-630 in cyanobacteria 640, 644-649 photosynthetic electron transport 31, 145 isolation 102 photosynthetic membrane biogenesis 632-639, LD spectrum 321 702 lipolytic hydrolase effects 724 photosynthetic membrane morphology 1-72, localization in thylakoids 31, 34-38, 517, 146-158, 603-618, 620-630 671-672 photosynthetic unit 288, 633 mobility 521 photosynthetic water oxidation 437-445, mutants 34, 671-672 447-455 oxygen-evolving preparations 38 photosystem I (see also P-700), assembly 492, oxygen evolving system 437-445, 447-455 493 polypeptides 433, 434 chlorophyll-proteins 101, 103, 108, 111, 275 populations 335 complexes 39, 40, 109-112, 160, 168-172, primary reactants 422

492 QB protein 380, 457-463, 468-474 core complex 40, 169 reaction center 422-435, 457 cyanobacteria 118, 640, 647, 649, 650 reaction centers, bacteria vs. plants digitonin particles 553 371-380, 383, 457 energy conversion 87 reconstituted complexes 38 fluorescence quenching 313 redox potentials 426, 427 LD peak 321 relationship to phycobilisomes 56-58, 226, lipolytic hydrolase effects 724 331 localization on fracture faces 39, 40, 492, siphonaxanthin-chl a/b proteins 275 669 subunits 457, 458 localization in thylakoids 35, 39, 40, 517, photosystem II to photosystem I ratio 53, 517 669-671 photosystem II, 34 kDa protein 453 mobility 521 47 kDa protein 380, 453, 596 of algae 273 a-centers 37, 38, 304, 433 of Prochloron 276 a-ß heterogeneity 308 reaction center electron transfer 476-485, photosystem II, ^-centers 37, 38 553 phycobilin 244 reconstitution 39, 679 phycobiliproteins (see also phycobilisomes) relationship to phycobilisomes 266 117, 118, 241, 260-268, 327 similarities with bacterial reaction centers heterodimers 250 208, 371 phycobilisomes 260-268 siphoxanthin-chl a/b proteins 276 association with photosystem II 56-58, subunit III 553-554 266-269, 644 surface charge 552 compared to chlorosomes 393 Triton X-100 particles 553 emission 262 photosystem II 87 hemispherical type 26 antennae 112-115, 264 isolation 261 bound antennae 44 light regulation 18 carotenoid oxidation 430 linker peptides 329 charge separation 422 molecular ecology 328 chlorophyll a-proteins 273 polypeptides 241, 331 chlorophyll-proteins 101, 103, 108 reconstitution 266 Cl~ requirement 573 structure 18, 53, 262-266, 329 cofactors 457 phycocyanin 241, 280 composition 112-115,158-168, 457-467 phycocyanobilin 327 core complex 35, 458 phycoerythrin 241, 280 cytochrome b-559 430 phycoerythrobilin 327 800 Subject Index

phycourobilin 327 Prosthecochloris 127 phylogenetic relationship of pigment proteins protease digestion of L and M subunits 622 244, 248 protein acid-base dissociation groups 570 phytochrome 330 protein synthesis associated with photoinhibi- phytoplankton 469 tion 473, 474 phytyl side chains 375 protein targeting 638 phytylation 690 prothylakoids 70, 686, 687 picosecond fluorescence decays 299 protochlorophyllide reductase 687 picosecond laser pulse 310 proton buffering domains 571 picosecond time-resolved techniques 333 proton channel forming proteins 574 pigment arrangement 374 proton currents, intramembrane 567 terminal 264 proton gradient 573 pigment-proteins (see also light-harvesting com• formation kinetics 563 plexes), spectral analysis 99 proton pump 536, 537 synthesis 611 proton translocation 184, 185, 529, 530, 544, pigments, distribution among algae 16, 269 560, 570 Pisum 385 protonic circuits 567 plants, shade 24 protonmotive force 180, 184, 185 plastocyanin 145, 160, 168-179, 512, 518, PS I (see photosystem I) 547-559 PS II (see photosystem II)

binding site at PS I 554 PS IIa 37, 49, 304, 308, 433

chemical modification 550 PS llß 37, 49, 308

electron transfer site 550, 551 psbA gene for Dl (QB) protein 385, 463 inhibitors 550 psbD gene for D2 protein 463 isoelectric point (pi) 552 Pseudoanabaena 264 mobility 179, 518-524, 556 puddle model 312 molecular structure 171, 548, 549 pyrenoids 11-13 rate constants 552, 553 reaction with cytochrome f 555 synthesis 547 Qpool 218,541 plastocyanin to P-700 ratio 557 Q-cycle 174-176,534-537,542

plastoglobuli 1 Q2 432 plastohydroquinone 441, 724 Q400 432 plastoquinol 145, 173-176 QA 228,424,425,427 plastoquinone (see also quinone) 153-156, QB 379,433,457-462,471 172, 177, 518, 531 Qß-protein 162-165, 468-474, 596, 597 and cytochrome b6/f 174 herbicides 460 and photosystem II 158-164 mutations 462, 465 mobility 178, 179, 521-522, 695, 724 turnover 461, 468-474 plastosemiquinone 165, 429, 471 Qc 541 polarized emission 322 QL 431 polypeptides, primary sequence of light-harvest• quantum efficiency 162 ing 382, 240 quantum yield 338 Porphyridium 2, 26, 261, 331, 333, 334 quinol-cytochrome c oxidoreductases 208, 496 Portulacca 5 quinone (see also plastoquinone) 121, 122, post-translationally imported proteins 451 540, 541 potential, transmembrane 566 quinone binding 209,231,382,625 primary acceptor, of bacteria 354 quinone radical 471 of photosystem I 477 quinone-iron complex 358

of photosystem II 324 QZ 541 primary donor, exciton trapping 325 of photosystem I 477 primary reactions in bacteria 338 radical formation 595 primary structure 248 radical pair 339, 341 Prochloron 15, 19, 143, 276 RC-H polypeptide 636 Prochlorophyta 276 RC-L, organization of structural gene 633 prolamellar bodies 70-72, 683-692 RC-M, organization of structural gene 633 Subject Index 801 reaction center, biochemistry 127 S-states 166-168, 437 cytochrome c 220 SAN-9785 24 four types 371 Sargassum 60 general reaction scheme 205 Scenedesmus 685, 689 lake model 313 secondary electron transfer 355 electron transfer events 144,161 secondary quinone acceptor 354 generated membrane potential 203, 206 semi-aerobic induction 609 organization 85 sensitizers 331 to phycobilisome ratio 56-58, 267 sequence homology 248 topography 124, 125 signal II 165,166,170,428 reactions centers of bacteria 107, 205, 218, single site catalysis 589 354, 357, 371-380, 382, 621 single turnover resolution 564 of green bacteria 371, 394, single-photon timing 299, 333 reaction center I complex 488, 489 siphonaxanthin-chl a/b proteins 275 reaction center I composition 152, 487-494 siphonoxanthin 271, 272 reaction center I organization 168-170, slow component lifetimes 302 476-485 slow electrogenic phase 528-529 reaction center II composition 152, 457-466 sodium dodecyl sulfate 104, 129 reaction center II, charge separation 162, space charge density 654 422-434 special pair 360 manganese depletion 35, 167 Spermothamnion 26, 53 reaction center-bCi complex cycle 211 spillover 303 reaction center-phycobilisome coupling 56-58, spin axes 402, 406 267 spin polarization 483 reconstitution of membrane complexes 38, 39, spin-probes 724 613, 672, 675-681, 722 spinach 59, 153, 173, 183 redox center orientation in bacterial mem• spirilloxanthin 125 branes 346-348, 351 Spirodela 684 redox centers in reaction centers of bacteria staining, tetramethylbenzidine 641 344 State I-State II transitions 64-66, 115, 657, redox chain control 561 658 redox potentials, theory and conventions 204 steady-state coupling 560-561 reducing equivalents 219 Stern layer 654 reducing power sources 212 stress 472, 473 relocalization 286 stroma 10-14, 665 repair of photodamage 474 membranes 20-24, 147, 305 repetitive streak cameras 333 strong coupling 292, 333 resonance energy transfer 92 structure analysis, x-ray 328, 372 respiration in mitochondria 496 structure, bacterial reaction center 345 reverse electron flow 164, 530 cyt b/Ci complex 345 Rhodeila 26 primary 240, 242, 328, 348 Rhodophyceae 6 secondary 346, 351 Rhodophyta 143,270,281,282,284 three dimensional of proteins 240, 244 Rhodopseudomonas 118, 119, 121, 123, 125, topographical 504 197-232, 246, 254, 256, 313, 344, 355, 371- subchloroplast fractions 696 381, 383-389, 497, 539, 545, 604, 605, 610-618 surface charge of thylakoids 154-158, Rhodospirillum 99, 125, 219, 246, 253, 313, 653-664, 698, 700 471, 610-618 sulfide, electron transport to reaction center ribulose bisphosphate carboxylase 11 215, 216 Rieske FeS center 173, 174, 498, 499 sulfide oxidation 217 Rieske FeS protein 127, 500-502, 540, 649 sulfolipids (see diacylsulfoquinovosylglycerol) gene 503 sulfur compounds, reduced 215 structure 504-605 transport 220 rise time of fluorescence 306, 334 sulfur-oxidizing enzymes 218 rod elements of chlorosomes 610, 618 sulphoquinovosyldiacylglycerol (see diacylsulfo• rotary shadowed replicas 666 quinovosylglycerol) RYDMR 364-367 sunlight 86 802 Subject Index

superexchanges 358 stroma development 20-22 supramolecular organization of thylakoids stroma three dimensional organization 1-7, 603-618, 675-681 19-20, 63 model 49 surface, inner 451 surface potential 653 tightly-bound nucleotides 589-592 Synechococcus 17 time-resolved spectrum 305 Synechocystis 263 tobacco 3 systematic organization of algae 5 Tolypothrix 263 topology of antenna pigments 317 topology of membranes 281, 620-630 TADMR 401,410 transient coupling 563-564 temperature dependence of fluorescence 307 transient states 338, 342 temperature-induced changes in membranes transmembrane electrochemical potential 145, 66-70 170, 176, 180, 181, 184, 187 terminal polypeptide antibodies 265 transport, Mg+ + control 229 tertiary electron transfer 355 trapping of excitation energy 286, 293 tetranitromethane 551 triazines 164, 165, 595 tetraphenyl boron 431 triplet states 292, 293, 310, 340, 356, 363, 395, Thiocapsa 122 400-421, 482 thioether bonds 327 triplet-minus-singlet absorbance difference thioredoxin 186, 590 402, 409-412 thiosulfate oxidation 217 triplet-minus-singlet spectra 417-418 thylakoid, architecture 1-72, 513-516, Tris, alkaline release of proteins 448 603-618, 665-673 Triton X-100 106, 129 arrangement in chloroplasts 6, 14-30 tunneling of electrons 95

fragments 299, 305, 306 turnover of QB protein 471, 472 frets 515 two electron gate 163, 174 freeze-fractured 1-72, 603-618, 665-673 freshly prepared 573 ubiquinone 119, 178, 372 frozen and thawed 573 unstacking in vitro 61-63, 149, 660, 616 lipids 683-691, 693-704, 706-711, 713-724 urea 595

location of cyt b6/f 40 marginal regions 152, 722 nomenclature 2, 22-23 van der Waals attractive forces 657 of chromophytan algae 28-30 variable fluorescence yield 306 of Chrysophyceae 29 vesicles, inside-out 305, 448 of cryptomonads 28 right-side-out 448 of cyanelles 53-58 violaxanthin-chl a/cl/c2-protein 277 of cyanobacteria 53-58 violaxanthin-chl a/chl cl/c2 complex 278 of diatoms 28-29 violaxanlhin-chlorophyll a/cl +c2-complex of dinoflagellates 29-30 116 of euglenoids 25 vitamin C 601 of green algae 25 of oxygen-evolving prokaryotes 16-19 water oxidation site 453 of Prochloron 51-52 water stres effects on photoinhibition 468 of red algae 53-58 water-splitting enzymes 37, 447-455 of Xanthophyceae 29 supramolecular organization 30-61, 603-618, 665-673 X-ray analysis of proteins 330, 357, 371, 624 X 432 protein density 516-518 a thylakoids, brown algae 28-29 endogenous buffering capacity 563 Z 428,429 girdle 26 Z+ 165,429,440 grana development 20-22 Z-scheme 144, 145 grana three dimensional organization 19-22, Zea 23 146-149 zero field splitting parameters 403, 404 stacking, light intensity 23 zeta-potential 655