Characterization of a Mo-Nitrogenase Variant Containing a Citrate-Substituted Cofactor

Liedtke, Jasper; Lee, Chi Chung; Tanifuji, Kazuki; Jasniewski, Andrew J.; Ribbe, Markus W.; Hu, Yilin

doi:10.1002/cbic.202000598

Title: Characterization of a Mo-Nitrogenase Variant Containing a Citrate-Substituted Cofactor

Journal Article · Sat Sep 12 00:00:00 EDT 2020 · ChemBioChem: a European journal of chemical biology

DOI:https://doi.org/10.1002/cbic.202000598· OSTI ID:1785240

Liedtke, Jasper ^[1]; Lee, Chi Chung ^[1]; Tanifuji, Kazuki ^[1]; Jasniewski, Andrew J. ^[1];

^[1]; Hu, Yilin ^[1]

Univ. of California, Irvine, CA (United States)

Nitrogenase converts N₂ to NH₃, and CO to hydrocarbons, at its cofactor site. In this work, we report a biochemical and spectroscopic characterization of a Mo-nitrogenase variant expressed in an Azotobacter vinelandii strain containing a deletion of nifV, the gene encoding the homocitrate synthase. Designated NifDK^Cit, the catalytic component of this Mo-nitrogenase variant contains a citrate-substituted cofactor analogue. Activity analysis of NifDK^Cit reveals a shift of CO reduction from H₂ evolution toward hydrocarbon formation and an opposite shift of N₂ reduction from NH₃ formation toward H₂ evolution. Consistent with a shift in the Mo K-edge energy of NifDK^Cit relative to that of its wild-type counterpart, EPR analysis demonstrates a broadening of the line-shape and a decrease in the intensity of the cofactor-originated S=3/2 signal, suggesting a change in the spin properties of the cofactor upon citrate substitution. These observations point to a crucial role of homocitrate in substrate reduction by nitrogenase and the possibility to tune product profiles of nitrogenase reactions via organic ligand substitution.

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Research Organization:: Univ. of California, Irvine, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: SC0016510; SC0014470

OSTI ID:: 1785240

Journal Information:: ChemBioChem: a European journal of chemical biology, Vol. 22, Issue 1; ISSN 1439-4227

Publisher:: ChemPubSoc EuropeCopyright Statement

Country of Publication:: United States

Language:: English

References (27)

Mechanism of Molybdenum Nitrogenase Burgess, Barbara K.; Lowe, David J. Chemical Reviews, Vol. 96, Issue 7 https://doi.org/10.1021/cr950055x	journal	January 1996
Structural basis of biological nitrogen fixation Rees, Douglas C.; Akif Tezcan, F.; Haynes, Chad A. Philosophical Transactions of the Royal Society A: Mathematical, Physical and Engineering Sciences, Vol. 363, Issue 1829 https://doi.org/10.1098/rsta.2004.1539	journal	April 2005
Electron Transfer in Nitrogenase Rutledge, Hannah L.; Tezcan, F. Akif Chemical Reviews, Vol. 120, Issue 12 https://doi.org/10.1021/acs.chemrev.9b00663	journal	January 2020
Reactivity, Mechanism, and Assembly of the Alternative Nitrogenases Jasniewski, Andrew J.; Lee, Chi Chung; Ribbe, Markus W. Chemical Reviews, Vol. 120, Issue 12 https://doi.org/10.1021/acs.chemrev.9b00704	journal	March 2020
Reduction of Substrates by Nitrogenases Seefeldt, Lance C.; Yang, Zhi-Yong; Lukoyanov, Dmitriy A. Chemical Reviews, Vol. 120, Issue 12 https://doi.org/10.1021/acs.chemrev.9b00556	journal	March 2020
Vanadium Nitrogenase Reduces CO Lee, C. C.; Hu, Y.; Ribbe, M. W. Science, Vol. 329, Issue 5992 https://doi.org/10.1126/science.1191455	journal	August 2010
Extending the Carbon Chain: Hydrocarbon Formation Catalyzed by Vanadium/Molybdenum Nitrogenases Hu, Y.; Lee, C. C.; Ribbe, M. W. Science, Vol. 333, Issue 6043 https://doi.org/10.1126/science.1206883	journal	August 2011
A comprehensive mechanism for the Fischer-Tropsch synthesis Rofer-DePoorter, Cheryl K. Chemical Reviews, Vol. 81, Issue 5 https://doi.org/10.1021/cr00045a002	journal	October 1981
Tracing the Hydrogen Source of Hydrocarbons Formed by Vanadium Nitrogenase Lee, Chi Chung; Hu, Yilin; Ribbe, Markus W. Angewandte Chemie International Edition, Vol. 50, Issue 24 https://doi.org/10.1002/anie.201100869	journal	April 2011
The pathway for serial proton supply to the active site of nitrogenase: enhanced density functional modeling of the Grotthuss mechanism Dance, Ian Dalton Transactions, Vol. 44, Issue 41 https://doi.org/10.1039/C5DT03223G	journal	January 2015
The controlled relay of multiple protons required at the active site of nitrogenase Dance, Ian Dalton Transactions, Vol. 41, Issue 25 https://doi.org/10.1039/c2dt30518f	journal	January 2012
Catalytic Reduction of Hydrazine to Ammonia with MoFe ₃ S ₄ −Polycarboxylate Clusters. Possible Relevance Regarding the Function of the Molybdenum-Coordinated Homocitrate in Nitrogenase Demadis, Konstantinos D.; Malinak, Steven M.; Coucouvanis, Dimitri Inorganic Chemistry, Vol. 35, Issue 13 https://doi.org/10.1021/ic960098b	journal	January 1996
Purification of the Azotobacter vinelandii nifV-encoded homocitrate synthase. Zheng, L.; White, R. H.; Dean, D. R. Journal of Bacteriology, Vol. 179, Issue 18 https://doi.org/10.1128/jb.179.18.5963-5966.1997	journal	September 1997
Identification of the V factor needed for synthesis of the iron-molybdenum cofactor of nitrogenase as homocitrate Hoover, Timothy R.; Robertson, Andrew D.; Cerny, Ronald L. Nature, Vol. 329, Issue 6142 https://doi.org/10.1038/329855a0	journal	October 1987
Citrate substitutes for homocitrate in nitrogenase of a nifV mutant of Klebsiella pneumoniae Liang, Jihong; Madden, Mark; Shah, Vinod K. Biochemistry, Vol. 29, Issue 37 https://doi.org/10.1021/bi00489a011	journal	September 1990
Crystallographic Analysis of the MoFe Protein of Nitrogenase from a nifV Mutant of Klebsiella pneumoniae Identifies Citrate as a Ligand to the Molybdenum of Iron Molybdenum Cofactor (FeMoco) Mayer, Suzanne M.; Gormal, Carol A.; Smith, Barry E. Journal of Biological Chemistry, Vol. 277, Issue 38 https://doi.org/10.1074/jbc.M205888200	journal	July 2002
Requirement of nifV gene for production of wild-type nitrogenase enzyme in Klebsiella pneumoniae McLean, P. A.; Dixon, R. A. Nature, Vol. 292, Issue 5824 https://doi.org/10.1038/292655a0	journal	August 1981
Nitrogenase of Klebsiella pneumoniae nifV mutants. Investigation of the novel carbon monoxide-sensitivity of hydrogen evolution by the mutant enzyme McLean, P. A.; Smith, B. E.; Dixon, R. A. Biochemical Journal, Vol. 211, Issue 3 https://doi.org/10.1042/bj2110589	journal	June 1983
A V‐Nitrogenase Variant Containing a Citrate‐Substituted Cofactor Newcomb, Megan P.; Lee, Chi Chung; Tanifuji, Kazuki ChemBioChem, Vol. 21, Issue 12 https://doi.org/10.1002/cbic.201900654	journal	December 2019
The FeMoco-deficient MoFe Protein Produced by a nifH Deletion Strain of Azotobacter vinelandii Shows Unusual P-cluster Features Ribbe, Markus W.; Hu, Yilin; Guo, Maolin Journal of Biological Chemistry, Vol. 277, Issue 26 https://doi.org/10.1074/jbc.M202061200	journal	April 2002
Comparison of Iron-Molybdenum Cofactor-deficient Nitrogenase MoFe Proteins by X-ray Absorption Spectroscopy: IMPLICATIONS FOR P-CLUSTER BIOSYNTHESIS Corbett, Mary C.; Hu, Yilin; Naderi, Farzad Journal of Biological Chemistry, Vol. 279, Issue 27 https://doi.org/10.1074/jbc.M403156200	journal	April 2004
FeMo cofactor maturation on NifEN Hu, Y.; Corbett, M. C.; Fay, A. W. Proceedings of the National Academy of Sciences, Vol. 103, Issue 46 https://doi.org/10.1073/pnas.0602647103	journal	October 2006
Optimization of FeMoco Maturation on NifEN Yoshizawa, Janice M.; Blank, Michael A.; Fay, Aaron W. Journal of the American Chemical Society, Vol. 131, Issue 26 https://doi.org/10.1021/ja9035225	journal	July 2009
Structure of a Cofactor-Deficient Nitrogenase MoFe Protein Schmid, B. Science, Vol. 296, Issue 5566 https://doi.org/10.1126/science.1070010	journal	April 2002
Structural evidence for a dynamic metallocofactor during N ₂ reduction by Mo-nitrogenase Kang, Wonchull; Lee, Chi Chung; Jasniewski, Andrew J. Science, Vol. 368, Issue 6497 https://doi.org/10.1126/science.aaz6748	journal	June 2020
Catalysis-dependent selenium incorporation and migration in the nitrogenase active site iron-molybdenum cofactor Spatzal, Thomas; Perez, Kathryn A.; Howard, James B. eLife, Vol. 4 https://doi.org/10.7554/eLife.11620	journal	December 2015
Evidence for Interstitial Carbon in Nitrogenase FeMo Cofactor Spatzal, T.; Aksoyoglu, M.; Zhang, L. Science, Vol. 334, Issue 6058 https://doi.org/10.1126/science.1214025	journal	November 2011

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37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
ammonia
carbon monoxide
cofactors
hydrocarbons
molybdenum nitrogenase

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