Crystal Structure and Biochemical Characterization of Chlamydomonas FDX2 Reveal Two Residues that, When Mutated, Partially Confer FDX2 the Redox Potential and Catalytic Properties of FDX1

Boehm, Marko; Alahuhta, Markus; Mulder, David W.; Peden, Erin A.; Long, Hai; Brunecky, Roman; Lunin, Vladimir V.; King, Paul W.; Ghirardi, Maria L.; Dubini, Alexandra

doi:10.1007/s11120-015-0198-6

Title: Crystal Structure and Biochemical Characterization of Chlamydomonas FDX2 Reveal Two Residues that, When Mutated, Partially Confer FDX2 the Redox Potential and Catalytic Properties of FDX1

Journal Article · Tue Nov 03 00:00:00 EST 2015 · Photosynthesis Research

DOI:https://doi.org/10.1007/s11120-015-0198-6· OSTI ID:1242885

Boehm, Marko ^[1]; Alahuhta, Markus ^[1]; Mulder, David W. ^[1]; Peden, Erin A. ^[1]; Long, Hai ^[1]; Brunecky, Roman ^[1]; Lunin, Vladimir V. ^[1]; King, Paul W. ^[1]; Ghirardi, Maria L. ^[1]; Dubini, Alexandra ^[1]

National Renewable Energy Lab. (NREL), Golden, CO (United States)

The green alga Chlamydomonas reinhardtii contains six plastidic [2Fe2S]-cluster ferredoxins (FDXs), with FDX1 as the predominant isoform under photoautotrophic growth. FDX2 is highly similar to FDX1 and has been shown to interact with specific enzymes (such as nitrite reductase), as well as to share interactors with FDX1, such as the hydrogenases (HYDA), ferredoxin:NAD(P) reductase I (FNR1), and pyruvate:ferredoxin oxidoreductase (PFR1), albeit performing at low catalytic rates. Here we report the FDX2 crystal structure solved at 1.18 Å resolution. Based on differences between the Chlorella fusca FDX1 and C. reinhardtii FDX2 structures, we generated and purified point-mutated versions of the FDX2 protein and assayed them in vitro for their ability to catalyze hydrogen and NADPH photo-production. The data show that structural differences at two amino acid positions contribute to functional differences between FDX1 and FDX2, suggesting that FDX2 might have evolved from FDX1 toward a different physiological role in the cell. Moreover, we demonstrate that the mutations affect both the midpoint potentials of the FDX and kinetics of the FNR reaction, possibly due to altered binding between FDX and FNR. An effect on H₂ photo-production rates was also observed, although the kinetics of the reaction were not further characterized.

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Science (SC)

Grant/Contract Number:: AC36-08GO28308

OSTI ID:: 1242885

Report Number(s):: NREL/JA-2700-62499

Journal Information:: Photosynthesis Research, Vol. 128, Issue 1; Related Information: Photosynthesis Research; ISSN 0166-8595

Publisher:: SpringerCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 16 works

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References (50)

Pyruvate:Ferredoxin Oxidoreductase Is Coupled to Light-independent Hydrogen Production in Chlamydomonas reinhardtii Noth, Jens; Krawietz, Danuta; Hemschemeier, Anja Journal of Biological Chemistry, Vol. 288, Issue 6 https://doi.org/10.1074/jbc.M112.429985	journal	December 2012
Comparative studies on two ferredoxins from the cyanobacterium Nostoc strain MAC Hutson, K. G.; Rogers, L. J.; Haslett, B. G. Biochemical Journal, Vol. 172, Issue 3 https://doi.org/10.1042/bj1720465	journal	June 1978
The PsaC subunit of photosystem I provides an essential lysine residue for fast electron transfer to ferredoxin Fischer, N. The EMBO Journal, Vol. 17, Issue 4 https://doi.org/10.1093/emboj/17.4.849	journal	February 1998
An aromatic amino acid is required at position 65 in Anabaena ferredoxin for rapid electron transfer to ferredoxin:NADP+ reductase Hurley, John K.; Cheng, Hong; Xia, Bin Journal of the American Chemical Society, Vol. 115, Issue 25 https://doi.org/10.1021/ja00078a006	journal	December 1993
Overview of the CCP 4 suite and current developments Winn, Martyn D.; Ballard, Charles C.; Cowtan, Kevin D. Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4 https://doi.org/10.1107/S0907444910045749	journal	March 2011
The ferredoxin docking site of photosystem I Sétif, Pierre; Fischer, Nicolas; Lagoutte, Bernard Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1555, Issue 1-3 https://doi.org/10.1016/S0005-2728(02)00279-7	journal	September 2002
Atomic Resolution Modeling of the Ferredoxin:[FeFe] Hydrogenase Complex from Chlamydomonas reinhardtii Chang, Christopher H.; King, Paul W.; Ghirardi, Maria L. Biophysical Journal, Vol. 93, Issue 9 https://doi.org/10.1529/biophysj.107.108589	journal	November 2007
Higher Order Structure Contributes to Specific Differences in Redox Potential and Electron Transfer Efficiency of Root and Leaf Ferredoxins ^† Gou, Ping; Hanke, Guy T.; Kimata-Ariga, Yoko Biochemistry, Vol. 45, Issue 48 https://doi.org/10.1021/bi061779d	journal	December 2006
Crystal structure determination at 1.4 Å resolution of ferredoxin from the green alga Chlorella fusca Bes, M. Teresa; Parisini, Emilio; Inda, Luis A. Structure, Vol. 7, Issue 10 https://doi.org/10.1016/s0969-2126(00)80054-4	journal	October 1999
EasySpin, a comprehensive software package for spectral simulation and analysis in EPR Stoll, Stefan; Schweiger, Arthur Journal of Magnetic Resonance, Vol. 178, Issue 1, p. 42-55 https://doi.org/10.1016/j.jmr.2005.08.013	journal	January 2006
Chlamydomonas reinhardtii Chloroplasts Contain a Homodimeric Pyruvate:Ferredoxin Oxidoreductase That Functions with FDX1 van Lis, Robert; Baffert, Carole; Couté, Yohann Plant Physiology, Vol. 161, Issue 1 https://doi.org/10.1104/pp.112.208181	journal	November 2012
Multiple ferredoxin isoforms in Chlamydomonas reinhardtii – Their role under stress conditions and biotechnological implications Winkler, Martin; Hemschemeier, Anja; Jacobs, Jessica European Journal of Cell Biology, Vol. 89, Issue 12 https://doi.org/10.1016/j.ejcb.2010.06.018	journal	December 2010
Natural engineering principles of electron tunnelling in biological oxidation–reduction Page, Christopher C.; Moser, Christopher C.; Chen, Xiaoxi Nature, Vol. 402, Issue 6757 https://doi.org/10.1038/46972	journal	November 1999
Characterization of the Key Step for Light-driven Hydrogen Evolution in Green Algae Winkler, Martin; Kuhlgert, Sebastian; Hippler, Michael Journal of Biological Chemistry, Vol. 284, Issue 52 https://doi.org/10.1074/jbc.M109.053496	journal	October 2009
Amino Acid Sequence of the Major Component of Aphanothece sacrum Ferredoxin1 Hase, Toshiharu; Wada, Keishiro; Matsubara, Hiroshi The Journal of Biochemistry, Vol. 79, Issue 2 https://doi.org/10.1093/oxfordjournals.jbchem.a131076	journal	February 1976
A redox‐dependent interaction between two electron‐transfer partners involved in photosynthesis Morales, Renaud; Charon, Marie‐Hélène; Kachalova, Galina EMBO reports, Vol. 1, Issue 3 https://doi.org/10.1093/embo-reports/kvd057	journal	September 2000
Structure of the electron transfer complex between ferredoxin and ferredoxin-NADP⁺ reductase Kurisu, Genji; Kusunoki, Masami; Katoh, Etsuko Nature Structural Biology, Vol. 8, Issue 2, p. 117-121 https://doi.org/10.1038/84097	journal	February 2001
Secondary-structure matching (SSM), a new tool for fast protein structure alignment in three dimensions Krissinel, E.; Henrick, K. Acta Crystallographica Section D Biological Crystallography, Vol. 60, Issue 12 https://doi.org/10.1107/S0907444904026460	journal	November 2004
Guidelines for tunneling in enzymes Moser, Christopher C.; Anderson, J. L. Ross; Dutton, P. Leslie Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1797, Issue 9 https://doi.org/10.1016/j.bbabio.2010.04.441	journal	September 2010
Electron Transfer in Proteins Gray, Harry B.; Winkler, Jay R. Annual Review of Biochemistry, Vol. 65, Issue 1 https://doi.org/10.1146/annurev.bi.65.070196.002541	journal	June 1996
Impact of Mutations on the Midpoint Potential of the [4Fe-4S] ^+1,+2 Cluster and on Catalytic Activity in Electron Transfer Flavoprotein-ubiquinone Oxidoreductase (ETF-QO) ^† Usselman, Robert J.; Fielding, Alistair J.; Frerman, Frank E. Biochemistry, Vol. 47, Issue 1 https://doi.org/10.1021/bi701859s	journal	January 2008
Iron-Sulfur Proteins: Structure and Function Orme-Johnson, W. H. Annual Review of Biochemistry, Vol. 42, Issue 1 https://doi.org/10.1146/annurev.bi.42.070173.001111	journal	June 1973
Improved tools for biological sequence comparison. Pearson, W. R.; Lipman, D. J. Proceedings of the National Academy of Sciences, Vol. 85, Issue 8, p. 2444-2448 https://doi.org/10.1073/pnas.85.8.2444	journal	April 1988
Residue Glu-91 of Chlamydomonas reinhardtii ferredoxin is essential for electron transfer to ferredoxin-thioredoxin reductase Jacquot, Jean-Pierre; Stein, Mariana; Suzuki, Akira FEBS Letters, Vol. 400, Issue 3 https://doi.org/10.1016/s0014-5793(96)01407-x	journal	January 1997
Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency Kapust, Rachel B.; Tözsér, József; Fox, Jeffrey D. Protein Engineering, Design and Selection, Vol. 14, Issue 12 https://doi.org/10.1093/protein/14.12.993	journal	December 2001
Microalgae: a green source of renewable H2 Ghirardi, M. Trends in Biotechnology, Vol. 18, Issue 12 https://doi.org/10.1016/S0167-7799(00)01511-0	journal	December 2000
Structural analysis of interactions for complex formation between Ferredoxin-NADP+ reductase and its protein partners Mayoral, Tomás; Martínez-Júlvez, Marta; Pérez-Dorado, Inmaculada Proteins: Structure, Function, and Bioinformatics, Vol. 59, Issue 3 https://doi.org/10.1002/prot.20450	journal	March 2005
Critical Residues of Chlamydomonas reinhardtii Ferredoxin for Interaction with Nitrite Reductase and Glutamate Synthase Revealed by Site-Directed Mutagenesis Garcia-Sanchez, M. Isabel; Gotor, Cecilia; Jacquot, Jean-Pierre European Journal of Biochemistry, Vol. 250, Issue 2 https://doi.org/10.1111/j.1432-1033.1997.0364a.x	journal	December 1997
Structural Basis for the Thermostability of Ferredoxin from the Cyanobacterium Mastigocladus laminosus Fish, Alexander; Danieli, Tsafi; Ohad, Itzhak Journal of Molecular Biology, Vol. 350, Issue 3 https://doi.org/10.1016/j.jmb.2005.04.071	journal	July 2005
Accurate bond and angle parameters for X-ray protein structure refinement Engh, R. A.; Huber, R. Acta Crystallographica Section A Foundations of Crystallography, Vol. 47, Issue 4 https://doi.org/10.1107/S0108767391001071	journal	July 1991
Mapping of Protein-Protein Interaction Sites in the Plant-Type [2Fe-2S] Ferredoxin Kameda, Haruka; Hirabayashi, Kei; Wada, Kei PLoS ONE, Vol. 6, Issue 7 https://doi.org/10.1371/journal.pone.0021947	journal	July 2011
Identification of Global Ferredoxin Interaction Networks in Chlamydomonas reinhardtii Peden, Erin A.; Boehm, Marko; Mulder, David W. Journal of Biological Chemistry, Vol. 288, Issue 49 https://doi.org/10.1074/jbc.M113.483727	journal	October 2013
Genetic disruption of both Chlamydomonas reinhardtii [FeFe]-hydrogenases: Insight into the role of HYDA2 in H2 production Meuser, Jonathan E.; D’Adamo, Sarah; Jinkerson, Robert E. Biochemical and Biophysical Research Communications, Vol. 417, Issue 2 https://doi.org/10.1016/j.bbrc.2011.12.002	journal	January 2012
Structure of CyanoP at 2.8 Å: Implications for the Evolution and Function of the PsbP Subunit of Photosystem II, Michoux, Franck; Takasaka, Kenji; Boehm, Marko Biochemistry, Vol. 49, Issue 35 https://doi.org/10.1021/bi1011145	journal	September 2010
MolProbity : all-atom structure validation for macromolecular crystallography Chen, Vincent B.; Arendall, W. Bryan; Headd, Jeffrey J. Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 1 https://doi.org/10.1107/S0907444909042073	journal	December 2009
Pattern of Expression and Substrate Specificity of Chloroplast Ferredoxins from Chlamydomonas reinhardtii Terauchi, Aimee M.; Lu, Shu-Fen; Zaffagnini, Mirko Journal of Biological Chemistry, Vol. 284, Issue 38 https://doi.org/10.1074/jbc.M109.023622	journal	July 2009
Features and development of Coot Emsley, P.; Lohkamp, B.; Scott, W. G. Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 4 https://doi.org/10.1107/S0907444910007493	journal	March 2010
Amino acid residues in Anabaena ferredoxin crucial to interaction with ferredoxin-NADP+ reductase: Site-directed mutagenesis and laser flash photolysis Hurley, J. K.; Salamon, Z.; Meyer, T. E. Biochemistry, Vol. 32, Issue 36 https://doi.org/10.1021/bi00087a013	journal	September 1993
REFMAC 5 for the refinement of macromolecular crystal structures Murshudov, Garib N.; Skubák, Pavol; Lebedev, Andrey A. Acta Crystallographica Section D Biological Crystallography, Vol. 67, Issue 4 https://doi.org/10.1107/S0907444911001314	journal	March 2011
Interaction of ferredoxin with ferredoxin:NADP reductase: Effects of chemical modification of ferredoxin Vieira, Barbara J.; Davis, Danny J. Archives of Biochemistry and Biophysics, Vol. 247, Issue 1 https://doi.org/10.1016/0003-9861(86)90542-4	journal	May 1986
Biomolecular EPR Spectroscopy Hagen, Wilfred Raymond https://doi.org/10.1201/9781420059588	book	December 2008
Plant type ferredoxins and ferredoxin-dependent metabolism: Chloroplast ferredoxins Hanke, Guy; Mulo, Paula Plant, Cell & Environment, Vol. 36, Issue 6 https://doi.org/10.1111/pce.12046	journal	January 2013
Structure−Function Relationships in Anabaena Ferredoxin: Correlations between X-ray Crystal Structures, Reduction Potentials, and Rate Constants of Electron Transfer to Ferredoxin:NADP ⁺ Reductase for Site-Specific Ferredoxin Mutants ^† ^, ^‡ Hurley, John K.; Weber-Main, Anne M.; Stankovich, Marian T. Biochemistry, Vol. 36, Issue 37 https://doi.org/10.1021/bi9709001	journal	September 1997
Structure and Function of Plant-Type Ferredoxins Fukuyama, Keiichi Photosynthesis Research, Vol. 81, Issue 3 https://doi.org/10.1023/b:pres.0000036882.19322.0a	journal	January 2004
Ferredoxin and flavodoxin reduction by photosystem I Sétif, Pierre Biochimica et Biophysica Acta (BBA) - Bioenergetics, Vol. 1507, Issue 1-3 https://doi.org/10.1016/S0005-2728(01)00205-5	journal	October 2001
Browsing gene banks for Fe2S2 ferredoxins and structural modeling of 88 plant-type sequences: An analysis of fold and function Bertini, Ivano; Luchinat, Claudio; Provenzani, Alessandro Proteins: Structure, Function, and Genetics, Vol. 46, Issue 1 https://doi.org/10.1002/prot.10009	journal	December 2001
Optimized Expression and Purification for High-Activity Preparations of Algal [FeFe]-Hydrogenase Yacoby, Iftach; Tegler, Lotta Tollstoy; Pochekailov, Sergii PLoS ONE, Vol. 7, Issue 4 https://doi.org/10.1371/journal.pone.0035886	journal	April 2012
Molecular replacement with MOLREP Vagin, Alexei; Teplyakov, Alexei Acta Crystallographica Section D Biological Crystallography, Vol. 66, Issue 1 https://doi.org/10.1107/S0907444909042589	journal	December 2009
Ferredoxin from a Blue-green Alga, Aphanothece sacrum (Suringar) OKADA1 Wada, Keishiro; Kagamiyama, Hiroyuki; Shin, Masateru The Journal of Biochemistry, Vol. 76, Issue 6 https://doi.org/10.1093/oxfordjournals.jbchem.a130674	journal	December 1974
Spinach Ferredoxin Matsubara, H.; Sasaki, R. M. Journal of Biological Chemistry, Vol. 243, Issue 8 https://doi.org/10.1016/S0021-9258(18)93507-6	journal	April 1968

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Title: Crystal Structure and Biochemical Characterization of Chlamydomonas FDX2 Reveal Two Residues that, When Mutated, Partially Confer FDX2 the Redox Potential and Catalytic Properties of FDX1

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