Louisiana State University LSU Digital Commons LSU Historical Dissertations and Theses Graduate School 1977 Application of Electron Spin Resonance: I. Photosystem I in Green Plant Photosynthesis. II. Malignant Tumor Associated Hematological Changes. Anupam Das gupta Louisiana State University and Agricultural & Mechanical College Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_disstheses Recommended Citation Das gupta, Anupam, "Application of Electron Spin Resonance: I. Photosystem I in Green Plant Photosynthesis. II. Malignant Tumor Associated Hematological Changes." (1977). LSU Historical Dissertations and Theses. 3103. https://digitalcommons.lsu.edu/gradschool_disstheses/3103 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Historical Dissertations and Theses by an authorized administrator of LSU Digital Commons. For more information, please contact [email protected]. INFORMATION TO USERS This malarial was produoad from a microfilm copy of tha original documant. Whila the moat advanoad tachnological maant to photopaph and taproduoa this documant have baan used, tha quality ia heavily dapandant upon tha quality of tha original aubmittad. Tha following ax pi ana ti on of tachniquaa ia providad to halp you understand marfclnga or pattama which may appear on thia reproduction. 1. Tha aign or "target" for pagaa apporandy lacking from tha documant photographed ia "Miaaing Paga(a)". If it waa poaaibla to obtain tha miaaing page(i) or taction, they are apliead into tha film along with adjacent pagaa. Thia may hava nacaasitatad cutting thru an image and duplicating adjaoant pagaa to inaura you complete continuity. 2. Whan an imap on the film ia obliterated with a large round black mark, it is an indication that tha photopaphar auapactad that tha copy may hava moved during exposure and thus causa a blurred image. You will find a good image of tha page in tha adjaoant frame. 3. Whan a map, drawing or chart, etc., waa part of tha material being photographed die photographer followed a definite method in "sectioning" tha material. It is customary to begin photoing at tha upper left hand corner of a large dieet and to continue photoing from left to right in equal sections with a small overlap. If nacaasary, sectioning is continued again - beginning below the first row and continuing on until complete. 4. Tha majority of ueers indicate that tha textual content is of greatest value, however, a somewhat higher quality reproduction could be made from "photographs" if essential to tha understanding of tha dissertation. Silver prints of "photographs" may be ordered at additional charge by writing tha Order Department, giving tha catalog number, tide, author and specific pages you wish reproduced. 5. PLEASE NOTE: Soma pages may hava indistinct print. Filmed as received. University Microfilms International 300 North Zaab Road Ann Arbor. Michigan 40106 USA St. John's Road. Tytor's Grosn High Wycombs. Bucks. England HP10 8HR 77-28,668 DAS GUPTA. Anupam, 1946- APPLICATION OF ELECTRON SPIN RESONANCE: I. PHOTOSYSTEM I IN GREEN PLANT PHOTO­ SYNTHESIS. II, MALIGNANT TUMOR ASSOCI­ ATED HEMATOLOGICAL CHANGES. Louisiana State University and Agricultural and Mechanical College, Ph.D., 1977 Biophysics, general Xerox University Microfilms , Ann Arbor. Michigan 40106 APPLICATION OP ELECTRON SPIN RESONANCE: PHOTOSYSTEM I IN GREEN PLANT PHOTOSYNTHESIS MALIGNANT TUMOR ASSOCIATED HEMATOLOGICAL CHANGES A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in The Department of Physics and Astronomy by Anupam Das Gupta B.S., Gauhati University, 1965 M.Sc., Calcutta University, 1968 M.S., Louisiana State University, 1972 August, 1977 EXAMINATION AND THESIS REPORT Candidate: Anupam Da a Gupta Major Field: Physics Title of Thetis: Application of Electron Spin Reaonance: I. Photoayatem 1 in Green Plant Photoayntheala II. Malignant Tumor Aaaociated Hematological Changea Approved: M ajor f Dean of the Graduate school EXAMINING COMMITTEE: Date of Examination: December 1, 1976 ACKNOWLEDGEMENT I wish to express my sincere appreciation to Prof. William 0. Hamilton and Prof, Brian J. Hales, under whose direction this work was performed, for their guidance, continuous encouragement, inspiration and advice which made the completion of this work possible. Financial assistance pertinent to the preparation of this dissertation was provided by the Charles E. Coates Memorial Fund of Louisiana State University donated by George H. Coates. I gratefully acknowledge this support and assistance. I would like to thank Mr. L, Edelen for tremendous help with the electronics, Ms. Nancy Harris for the drawings and Ms. Jackie Tamas for an excellent Job of typing. Finally I would like to thank my parents Mr, and Mrs. Anil Bhusan Das Gupta and my wife Rumu for their sacrifice, patience, understanding and encouragement during the years of my graduate work. ii DEDICATION To My Parents Anil Bhusan Das Gupta and Dipall Das Gupta and to My Grandmothers Sarojbala Das Gupta and Pritilata Kar 111 TABLE OP CONTENTS Page ACKNOWLEDGEMENT.......................... ii DEDICATION................................................ill LIST OF TABLES........................................... vli LIST OP FIGURES........................................ viii ABSTRACT................................................. xiii PART I CHAPTER I. THEORY.......................................... 1 Introduction ................................... 1 Zeeman Interaction ............................. 1 Saturation ..................................... 3 Relaxation ..................................... 4 Line Broadening................................. 6 Bloch Equations................................. 8 Line Shapes........................................ 10 Spin Hamiltonian..................................16 g Tensor.......................................... 19 II. PHOTOSYNTHESIS....................................21 Background........................................ 21 Chloroplast Structure............................. 23 Photosynthetic Pigments........................... 25 Absorption Spectra ............................. 28 Photosynthetic Unit............................... 28 Evidence for the Existence of Two Photosystems ................................. 30 Oxidation Reduction Reaction .................. 31 Z Scheme.......................................... 35 ESR Study of the Two Photosystems................40 Fractionation...................................... 47 Scope of the Present Work.........................51 III. MATERIALS AND METHODS............................. 52 Extraction of Chloroplast.........................52 Ultrasonication....................................52 iv V TABLE OP CONTENTS (continued) CHAPTER Page Batch AUT Treatment............................... 53 Dialysis ............... 54 Column Electrophoresis........................... 5** SDS-Acrylamide Gel Electrophoresis ........... 62 ESR Measurement....................................68 Kinetic Study......................................68 Triggering Device..................................69 Liquid Helium Study............................... 73 IV. RESULTS AND DISCUSSION............................75 Triton X-100 ................................... 76 Triton X-100 and S D S ............................. 89 LDAO ...............................................96 ESR Experiment....................................98 Low Temperature ESR Study........................Ill Conclusion....................................... 118 Future Experiment.................................120 REFERENCES.............................................. 123 PART II CHAPTER I. INTRODUCTION......................................129 II. IRON METABOLISM................................... 137 III. EXPERIMENTAL METHOD............................... 142 Sample Collection.................................142 ESR Setup......................................... 142 Preliminary Findings............................ 1*43 Control Experiment ............................. 146 Computer Analysis.................................147 IV. RESULTS AND DISCUSSION...........................165 Future Experiment.................................172 REFERENCES.............................................. 174 SELECTED BIBLIOGRAPHY ................................. 177 Vi TABLE OP CONTENTS (continued) APPENDICES Page I. REACTION RATE CONSTANT........................ 178 II. DETERMINATION OFCHLOROPHYLL ...................180 VITA..................................................... 182 LIST OP TABLES PART I Page Table I. Amount of Dense and Less Dense Solutions for Preparing the Density Gradient. 60 PART II Table I. ESR Parameters Obtained by Computer Fit to Experimental D a t a ........................ 157 vii PART I LIST OP FIGURES Figure Page 1. Lorentzian and Gaussian absorption curves. 13 2. Lorentzian and Gaussian absorption first derivative curve ................................. 14 3- Molecular structure of Chi a and Chi b ............ 26 4. Absorption spectra of Chi a and Chi b.............. 29 5. Bacterial photosynthetic apparatus .............. 36 6 . Green plant photosynthetic apparatus ............ 37 7. Apparatus for column electrophoresis ...... 57 8 . Apparatus for gel electrophoresis...................65 9. Schematics