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Title: Cysteine as a ligand platform in the biosynthesis of the FeFe hydrogenase H cluster

Abstract

Hydrogenases catalyze the redox interconversion of protons and H2, an important reaction for a number of metabolic processes and for solar fuel production. In FeFe hydrogenases, catalysis occurs at the H cluster, a metallocofactor comprising a [4Fe–4S]H subcluster coupled to a [2Fe]H subcluster bound by CO, CN–, and azadithiolate ligands. The [2Fe]H subcluster is assembled by the maturases HydE, HydF, and HydG. HydG is a member of the radical S-adenosyl-L-methionine family of enzymes that transforms Fe and L-tyrosine into an [Fe(CO)2(CN)] synthon that is incorporated into the H cluster. Though it is thought that the site of synthon formation in HydG is the “dangler” Fe of a [5Fe] cluster, many mechanistic aspects of this chemistry remain unresolved including the full ligand set of the synthon, how the dangler Fe initially binds to HydG, and how the synthon is released at the end of the reaction. In order to address these questions, we show in this paper that L-cysteine (Cys) binds the auxiliary [4Fe–4S] cluster of HydG and further chelates the dangler Fe. We demonstrate that a [4Fe–4S]aux[CN] species is generated during HydG catalysis, a process that entails the loss of Cys and the [Fe(CO)2(CN)] fragment; on this basis, we suggestmore » that Cys likely completes the coordination sphere of the synthon. Finally, through spectroscopic analysis of HydG before and after the synthon is formed, we conclude that Cys serves as the ligand platform on which the synthon is built and plays a role in both Fe2+ binding and synthon release.« less

Authors:
 [1];  [2];  [2];  [1];  [1];  [1];  [3];  [1]
  1. Department of Chemistry, University of California, Davis, CA 95616,
  2. Department of Chemical Engineering, Stanford University, Stanford, CA 94305,
  3. Department of Chemical Engineering, Stanford University, Stanford, CA 94305,, Department of Bioengineering, Stanford University, Stanford, CA 94305
Publication Date:
Research Org.:
Stanford Univ., CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES); National Institutes of Health (NIH)
OSTI Identifier:
1235132
Alternate Identifier(s):
OSTI ID: 1356198
Grant/Contract Number:  
FG02-09ER46632; SC0002010; GM111025; GM104543; GM65440
Resource Type:
Published Article
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Volume: 112 Journal Issue: 37; Journal ID: ISSN 0027-8424
Publisher:
Proceedings of the National Academy of Sciences
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; 59 BASIC BIOLOGICAL SCIENCES; FeFe hydrogenase; metallocofactor biosynthesis; HydG

Citation Formats

Suess, Daniel L. M., Bürstel, Ingmar, De La Paz, Liliana, Kuchenreuther, Jon M., Pham, Cindy C., Cramer, Stephen P., Swartz, James R., and Britt, R. David. Cysteine as a ligand platform in the biosynthesis of the FeFe hydrogenase H cluster. United States: N. p., 2015. Web. doi:10.1073/pnas.1508440112.
Suess, Daniel L. M., Bürstel, Ingmar, De La Paz, Liliana, Kuchenreuther, Jon M., Pham, Cindy C., Cramer, Stephen P., Swartz, James R., & Britt, R. David. Cysteine as a ligand platform in the biosynthesis of the FeFe hydrogenase H cluster. United States. https://doi.org/10.1073/pnas.1508440112
Suess, Daniel L. M., Bürstel, Ingmar, De La Paz, Liliana, Kuchenreuther, Jon M., Pham, Cindy C., Cramer, Stephen P., Swartz, James R., and Britt, R. David. Mon . "Cysteine as a ligand platform in the biosynthesis of the FeFe hydrogenase H cluster". United States. https://doi.org/10.1073/pnas.1508440112.
@article{osti_1235132,
title = {Cysteine as a ligand platform in the biosynthesis of the FeFe hydrogenase H cluster},
author = {Suess, Daniel L. M. and Bürstel, Ingmar and De La Paz, Liliana and Kuchenreuther, Jon M. and Pham, Cindy C. and Cramer, Stephen P. and Swartz, James R. and Britt, R. David},
abstractNote = {Hydrogenases catalyze the redox interconversion of protons and H2, an important reaction for a number of metabolic processes and for solar fuel production. In FeFe hydrogenases, catalysis occurs at the H cluster, a metallocofactor comprising a [4Fe–4S]H subcluster coupled to a [2Fe]H subcluster bound by CO, CN–, and azadithiolate ligands. The [2Fe]H subcluster is assembled by the maturases HydE, HydF, and HydG. HydG is a member of the radical S-adenosyl-L-methionine family of enzymes that transforms Fe and L-tyrosine into an [Fe(CO)2(CN)] synthon that is incorporated into the H cluster. Though it is thought that the site of synthon formation in HydG is the “dangler” Fe of a [5Fe] cluster, many mechanistic aspects of this chemistry remain unresolved including the full ligand set of the synthon, how the dangler Fe initially binds to HydG, and how the synthon is released at the end of the reaction. In order to address these questions, we show in this paper that L-cysteine (Cys) binds the auxiliary [4Fe–4S] cluster of HydG and further chelates the dangler Fe. We demonstrate that a [4Fe–4S]aux[CN] species is generated during HydG catalysis, a process that entails the loss of Cys and the [Fe(CO)2(CN)] fragment; on this basis, we suggest that Cys likely completes the coordination sphere of the synthon. Finally, through spectroscopic analysis of HydG before and after the synthon is formed, we conclude that Cys serves as the ligand platform on which the synthon is built and plays a role in both Fe2+ binding and synthon release.},
doi = {10.1073/pnas.1508440112},
journal = {Proceedings of the National Academy of Sciences of the United States of America},
number = 37,
volume = 112,
place = {United States},
year = {Mon Aug 31 00:00:00 EDT 2015},
month = {Mon Aug 31 00:00:00 EDT 2015}
}

Journal Article:
Free Publicly Available Full Text
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https://doi.org/10.1073/pnas.1508440112

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Cited by: 43 works
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