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The Hill Reaction: in Vitro and in Vivo Studies
Chapter 7 The Hill Reaction: In Vitro and In Vivo Studies Edward A. Funkhouser1 and Donna E. Balint2 1Department of Biochemistry and Biophysics Texas A&M University College Station, Texas 77843-2128 2Department of Horticulture Texas A&M University College Station, Texas 77843-2133 Edward is a professor of biochemistry and biophysics and of plant physiology. He currently serves as associate head for undergraduate education. He received his B.S. in horticulture from Delaware Valley College in 1967 and his M.S. and Ph.D. from Rutgers University in plant physiology in 1969 and 1972, respectively. His research interests deal with the molecular responses of perennial plants to water-deficit stress. He received the Association of Former Students Distinguished Achievement Award for Teaching which is the University's highest award. He also received the Diversity Award from the Department of Multicultural Services, Texas A&M University. Donna is a graduate student in plant physiology. She has supervised the laboratories which accompany the introductory course in plant physiology. She received her B.S. from the University of Connecticut in 1984 and M.Sc. from Texas A&M in 1993. Her research interests involve studies of salinity stress in plants. She received the Association of Former Students Graduate Teaching Award which recognized her excellence in teaching. Reprinted from: Funkhouser, E. A. and D. E. Balint. 1994. The Hill reaction: In vitro and in vivo studies. Pages 109- 118, in Tested studies for laboratory teaching, Volume 15 (C. A. Goldman, Editor). Proceedings of the 15th Workshop/Conference of the Association for Biology Laboratory Education (ABLE), 390 pages. -
Forty Years of Microbial Photosynthesis Research: Where It Came from and What It Led to ∗
R. Clinton Fuller Photosynthesis Research 62: 1–29, 1999. 3 © 1999 Kluwer Academic Publishers. Printed in the Netherlands. Personal perspective Forty years of microbial photosynthesis research: Where it came from and what it led to ∗ R. Clinton Fuller Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst, MA 01003, USA Received 14 May 1999; accepted in revised form 15 August 1999 Key words: chloroplast, chlorosome, chromatophore, granules, inositol, Neurospora, path of carbon, pho- tosynthesis, polythdroxyalkanoate (PHA), prokaryote cellular inclusions, protozoan biochemistry, ribulose 1,5-bis-phosphate, Tetrahymena Abstract What follows is a very personal account of my professional life and the early years that preceded it. I have described the social and economic conditions in America and how the nineteen twenties and thirties nurtured our scientific future. The description of the early part of post-World War II research covers my experience in the areas of nutritional biochemistry, biochemical genetics and proceeds to photosynthesis. The latter era lasted around 35 years. For me the most memorable research accomplishments in which I was a participant during this period was the first demonstration of the primary carboxylation enzyme in an in vitro system in algal and higher plants as well to show that it was structurally associated with the chloroplast.Our group while at Oak Ridge and the University of Massachusetts assembled data that described the complete macromolecular assembly of the photosynthetic -
Photosynthesis and the Hill Reaction
Edexcel A Biology A-Level Core Practical 11 Investigate photosynthesis using isolated chloroplasts (the Hill reaction). www.pmt.education Dehydrogenase is an enzyme found in plant chloroplasts that is crucial to the light dependent stage of photosynthesis. In the light dependent stage, electrons are accepted by NADP. This reaction was discovered in 1938 by Robin Hill and thus is often called the Hill reaction. Dehydrogenase catalyses this reaction. When a redox indicator dye is present, such as DCPIP (which turns from blue to colourless when it is reduced), electrons are accepted by this instead. Equipment ● Leaf sample ● Scissors ● Mortar and pestle (cold) ● Nylon mesh ● Filter funnel ● Centrifuge ● Centrifuge tubes ● Ice-water-salt bath ● Glass rod ● Measuring cylinder ● Beaker ● Pipettes ● Bench lamp ● Buffer ● Isolation medium ● DCPIP solution Method 1. Remove stalks from leaf samples. Cut into small sections. Grind sample using a pestle and mortar and place into a chilled isolation solution. 2. Place several layers of muslin cloth into funnel and wet with isolation medium to filter sample into a beaker. 3. Suspend the beaker in an ice water bath to keep sample chilled. 4. Transfer to centrifuge tubes and centrifuge at high speed for 10 minutes. This will separate chloroplasts into the pellet. 5. Remove supernatant and add pellet to fresh isolation medium. 6. Store isolation solution on ice. 7. Set the colorimeter to the red filter. Zero using a cuvette containing chloroplast extract and distilled water. www.pmt.education 8. Place test tube in rack 30cm from light source and add DCPIP. Immediately take a sample and add to cuvette. -
An Appreciation of Robert Hill and His Reaction
Photosynthesis Research 73: 51–54, 2002. 51 © 2002 Kluwer Academic Publishers. Printed in the Netherlands. Atribute ‘And whose bright presence’ – an appreciation of Robert Hill and his reaction David Alan Walker University of Sheffield, 6, Biddlestone Village, Northumberland NE65 7DT, UK (e-mail:d.a.walker@sheffield.ac.uk) Received 15 August 2001; accepted in revised form 14 September 2001 Key words: chloroplasts, electron transport oxygen evolution, Robert Hill, methaemoglobin, oxidants Abstract The Hill reaction, its elucidation, and significance is briefly described. Hill oxidants, the role of the methemoglobin reducing factor and its relation to ferredoxin, and the part played by chloroplast envelopes are discussed. Reputedly the best multiple pun ever (flavored, as it was, with literary allusion) came, on an occasion in the last century, when Lord Maughn gave a gold coin to a boy who had helped him with his luggage. The boy was called Hill. This prompted an erudite bystander to declare ‘Hail smiling morn that tips the hills with gold.’ There is a later line in this same verse which reads ‘and whose bright presence darkness drives away.’ As in every sort of science, our understanding of photosynthesis has grown by the combined contri- butions, large and small, from researchers in every land. It is fair to say, however, that Robert (Robin) Hill’s ‘bright presence’ drove away a deal of darkness in the field of photosynthetic electron transport. His experiments (Hill 1965; Bendall 1994) in photosyn- thesis, which were to influence our thinking for 60 years or more (see e.g. Rich 1992), started, in pre- war Cambridge (Hill 1937, 1939) with what inevitably came to be known as ‘the Hill reaction.’ Figure 1. -
Three Overlooked Photosynthesis Papers of Otto Warburg
Photosynthesis Research https://doi.org/10.1007/s11120-021-00858-8 LETTER TO THE EDITOR Three overlooked photosynthesis papers of Otto Warburg (1883–1970), published in the 1940s in German and in Russian, on light‑driven water oxidation coupled to benzoquinone reduction Holger Dau1 · Boris Ivanov2 · Dmitry Shevela3 · William H. Armstrong4 · Govindjee Govindjee5 Received: 26 May 2021 / Accepted: 14 June 2021 © The Author(s), under exclusive licence to Springer Nature B.V. 2021 Abstract After a brief background on Otto Heinrich Warburg (1883–1970), and some of his selected research, we provide highlights, in English, of three of his papers in the 1940s—unknown to many as they were not originally published in English. They are: two brief reports on Photosynthesis, with Wilhelm Lüttgens, originally published in German, in 1944: ‘Experiment on assimilation of carbonic acid’; and ‘Further experiments on carbon dioxide assimilation’. This is followed by a regular paper, originally published in Russian, in 1946: ‘The photochemical reduction of quinone in green granules’. Since the 1944 reports discussed here are very short, their translations are included in the Appendix, but that of the 1946 paper is provided in the Supplementary Material. In all three reports, Warburg provides the frst evidence for and elaborates on light-driven water oxidation coupled to reduction of added benzoquinone. These largely overlooked studies of Warburg are in stark contrast to Warburg’s well-known error in assigning the origin of the photosynthetically formed dioxygen to carbonate. “Truth is more likely to come out of error if it is clear and defnite, than out of confusion, and my experience teaches me that it is better to hold an understood and intelligible opinion, even if it should turn out to be wrong, than to be content with a muddleheaded mixture * of conficting views, sometimes called Govindjee Govindjee impartiality, and often no better than no [email protected] opinion at all.” Holger Dau Otto Warburg [email protected] <https:// www. -
The Hill Reaction of Photosynthesis in Isolated Chloroplasts a Quantitative Approach
:How~T-.TDo-It- I:. The Hil Reaction of Photosynthesis in Isolated Chioroplasts A Quantitative Approach Rob L. Dean Downloaded from http://online.ucpress.edu/abt/article-pdf/58/5/303/47660/4450156.pdf by guest on 02 October 2021 The assay to determine the rate of + 2H20 - 4H 4e + 02 Stock Solutions photoreduction of 2,6-dichloropheno- lindophenol (DCPIP) described here Variants of this assay are widely 1. Chloroplast isolation buffer has been used in a new course entitled used in student laboratories but in the (modified from Hopkins 1990): "Scientific Methods in Biology." The published and unpublished laboratory 50 mM tricine, 400 mM sorbitol, purpose of this laboratory course is to manuals that I have encountered, the 10 mM NaCl, 2.5 mM MgCl2- acquaint students with experimental considerable potential for the quanti- 6H20, 1.25 mM MnCl2, 0.3 mM design and teach them how to evalu- tative analysis of the data that it pro- Na2EDTA. Adjust pH to 7.8 with ate and communicate their results. vides is not fully exploited. By refer- NaOH. One goal is to enhance the scientific ence to a standard curve of absorbance 2. Chloroplast reaction buffer (5X literacy of our students by integrating at 600 nm against DCPIP concentra- concentrated, modified from basic chemistry and arithmetic, used tion, one can determine the change in Hopkins 1990): routinely in research laboratories, into molarity and hence the number of 250 mM tricine, 500 mM sorbitol, their practical experience. moles of DCPIP reduced in a given 100 mM KCl, 25 mM MgCl2. The principles on which the assay is time. -
Obituary Essays in Biochemistry David Alan Walker (1928–2012) Volume 48 Epigenetics, Disease His Phd
Obituary ESSAYS IN BIOCHEMISTRY David Alan Walker (1928–2012) VOLUME 48 Epigenetics, Disease his PhD. This first, rather fierce, encounter with corner of the globe, particularly from Germany, Robin fired a lifetime collaboration and friendship Australia and the USA. Visitors flocked to the between these two giants of photosynthesis RHI. Working together with Tom Delieu, who and Behaviour research. He then took up a fellowship with had also moved to Sheffield, he developed the first Robin in an even smaller laboratory in prototype oxygen electrodes. The development Cambridge, working on photophosphorylation and commercialization of equipment became a contemporaneously with Doug Graham, Dan cornerstone of the RHI, alongside systems for the Arnon and Bob Whatley, and with Frederick simultaneous measurement of photosynthesis Members Sanger across the corridor. He was particularly gas exchange, chlorophyll fluorescence andDownloaded from http://portlandpress.com/biochemist/article-pdf/34/5/57/6360/bio034050057.pdf by guest on 26 September 2021 impressed to be asked to give a lecture in the other key parameters, including the leaf disc Botany School by F.F. Blackman. In 1958, he oxygen electrode. Equipment development was save 25% met Charles Whittingham, who offered him something that suited his perfectionist streak. a lectureship at Queen Mary College in the David truly believed that knowledge was a University of London. It was here that he met precious gift to be shared and enjoyed. By his own only £16.46 Tom Delieu, an exceptionally talented technician, admission, lecturing filled David with horror, and he supervised his first PhD student, Geoffrey but paradoxically he revelled in the process of RRP £21.95 Hind. -
Photosynthesis: I Course: Plant Physiology and Biochemistry (M.Sc)
Photosynthesis: I Course: Plant Physiology and Biochemistry (M.Sc) Pratibha Singh Department of Botany Photosynthesis is a process by which some unique living organism converts light energy in to chemical energy. The light is used to produce reducing equivalents (NADPH) and these reducing equivalents are used in the process of reduction of CO2 to sugars. : algae, blue green algae, plants, sulfur bacteria (1.1) Here H2A is electron donor CO2 is electron acceptor H2A =H2O in oxygenic (Oxygen releasing) photosynthetic organism; algae and plants H2A=H2S in anoxygenic photosynthetic bacteria; purple sulfur bacteria Equation No. 1.1 was given by C.V Neil upon finding that in some bacteria H2S is used as substrate rather than H2O *why oxygen and sulfur?? as both belong to the same group in periodic table, however oxidation of H2S will release lesser energy as compared to H2O as sulfur is more electrpositive and donate electron easily as compared to oxygen. Equation 1.1. also suggest that oxygen evolved by oxygenic photosynthetic bacteria comes from Water molecule and not from CO2 This was further shown by Robert Hill (the scientist who was also involved in studying oxygen binding activity of hemoglobin )in 1937 Hill reaction: 2 H2O + 2 A + (light, chloroplasts) → 2 AH2 + O2 A: electron acceptor 1. Chloroplast isolated 2. Chloroplast treated with light 3. Recorded oxygen evolution using hemoglobin. Hill harnessed the change in spectral property of hemoglobin upon oxygen binding as a measure of oxygen evolution 4. Amount of oxygen released was higher in the presence and absence of artificial electron acceptor (ferric oxalate salts) Conclusion: Photosynthetic cells only evolve oxygen in light when in the presence of extracts of leaves or certain ferric salts, and do not evolve oxygen from carbon dioxide. -
NIH-Record-2000-04-18.Pdf
B ~ C 0 R a Still The Second Best Thing About Payday Katz Sees Directorship as H I G H L I G H ,T., S Campus' Major Unseen Construction Chance to Make a Difference By Robert Bock Project-'NIH Business System' NIH Business By Rich McManus " I viewed it as a tremendous challenge," Community Plans said NIAMS director Dr. Stephen Katz about New System here is a large construction project on campus that began accepting his institute's direct orship in last September and will likely last for the next 5 years, 1995. "It was an opportunity t o take all consuming the attention of hundreds of workers, costing an Human vs. Ape T that I had learned in science and medicine estimated $50 million over 5 years, and affecting the daily activi Differences Probed and make a greater impact than I ever had ties of at least 5,000 NIH'ers. Unlike the turned earth, diverted before." traffic and unsettled parking prompted by construction of the new Clinical Research Center and laboratory Bldgs. 50 and 40, this He came to NIAMS from the National STEP Plans Session project- the NIH Business System (NBS)-is going up without the Cancer Institute, where he served as chief On Addiction usual signs visible to the average employee. And unlike the new of the Dermatology Branch, a position he buildings, the new system will eventually affect everyone who continues to hold. An expert in cancerous works at NIH. and inflammatory disorders of the skin, NCI, Japan Society If you purchase a pipette, prepare travel papers, pay bills, loan Katz has demonstrated that skin is a critical Collaborate or move a piece of equipment, or seek the advice of an outside part of the immune system. -
Evolution of the Z-Scheme of Photosynthesis: a Perspective
Photosynth Res DOI 10.1007/s11120-016-0333-z HISTORY AND BIOGRAPHY Evolution of the Z-scheme of photosynthesis: a perspective Govindjee1 · Dmitriy Shevela2 · Lars Olof Björn3 Received: 7 November 2016 / Accepted: 29 December 2016 © Springer Science+Business Media Dordrecht 2017 Abstract The concept of the Z-scheme of oxygenic pho- reducing it; (b) Experimental discovery of the two light tosynthesis is in all the textbooks. However, its evolution reactions and two pigment systems and the Z-scheme of is not. We focus here mainly on some of the history of its photosynthesis: Robert Emerson’s discovery, in 1957, of biophysical aspects. We have arbitrarily divided here the enhancement in photosynthesis when two light beams (one 1941–2016 period into three sub-periods: (a) Origin of the in the far-red region, and the other of shorter wavelengths) concept of two light reactions: first hinted at, in 1941, by are given together than when given separately; and the James Franck and Karl Herzfeld; described and explained, 1960 scheme of Robin Hill & Fay Bendall; and (c) Evolu- in 1945, by Eugene Rabinowitch; and a clear hypoth- tion of the many versions of the Z-Scheme: Louis Duys- esis, given in 1956 by Rabinowitch, of the then available ens and Jan Amesz’s 1961 experiments on oxidation and cytochrome experiments: one light oxidizing it and another reduction of cytochrome f by two different wavelengths of light, followed by the work of many others for more than 50 years. Submitted for publication in honor of Nathan Nelson, a world leader in the field of photosynthesis, and of T. -
Newsletter University of Wisconsin-Madison
NEWSLETTER UNIVERSITY OF WISCONSIN-MADISON For friends of the Department of Biochemistry at the University of Wisconsin–Madison Table of Contents From the Chair .............................................................. 3 Department Alumnus: ................................................. 17 Biochemistry Phase II ................................................... 4 Honors and Awards...................................................... 18 Laurens Anderson at 90 ................................................. 5 Remembering Bob Burris ............................................ 20 New Faculty Profile ....................................................... 6 Gene Regulation .......................................................... 22 Biochemistry Scholars Program .................................... 8 Staff Departures ........................................................... 25 Where we are and are going .......................................... 9 Remembering Henry Lardy ........................................ 26 International programs .................................................... In Memoriam .............................................................. 27 Our Department in England ...................................... 10 Biochemistry Graduate Degrees .................................. 28 Our Department inUganda ....................................... 11 Biochemistry Advisor Graduate Degrees ..................... 29 Our Department in India ........................................... 12 From the Labs ............................................................ -
Biochemistry of Photosynthesis</Article-Title>
Book Reviews Botany the plant illustrations. The former are Related Processes -Rabinowitch, with useful to trained systematists, but it will each serving its purpose. On December be difficultfor the average reader to dis- 30, 1954, The Evening Star of Washing- MEDICALBOTANY: PLANTS AF- tinguish species using only these descrip- ton, D.C. headlined-"Photosynthesis FECTINGMAN'S HEALTH tions. Many of the plant illustrationsare Basis of Life Achieved by U.S. Scien- by Walter H. Lewis and Memory P.F. from John Gerards's "Herball" (1597) tists." Thus, much has gone before, and Elvin-Lewis. 1977. John Wiley and and, although intriguing,they are not the the current book makes you realize that Sons, Inc. (605 Third Avenue, New best representatives of the species in there is much yet to come. York 10006). 515 p. $27.50. Downloaded from http://online.ucpress.edu/abt/article-pdf/39/9/562/36074/4446100.pdf by guest on 30 September 2021 question; some did not reproduce well This book starts simply and concisely, Although there are excellent books and many are too sytlized. The range of leading the reader and building as it treating agriculture and evolutionary topics covered in the book is best illus- goes. An early discussion of chloroplasts features of cultivated plants, and other trated by the chapter headings: (1) is good but gives the impression that books which have dictionary-like lists of injurious plants-internal poisons, aller- there are only three forms or types of both tropical and temperate plants used gens, mutagens/teratogens; (2) remedial chloroplasts, those in bacteria, blue-green by man, there has not been available a plants used for cancer, nervous system, algae, and spinach.