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Title: CO-Bridged H-Cluster Intermediates in the Catalytic Mechanism of [FeFe]-Hydrogenase CaI

Abstract

The [FeFe]-hydrogenases ([FeFe] H2ases) catalyze reversible H2 activation at the H-cluster, which is composed of a [4Fe-4S]H subsite linked by a cysteine thiolate to a bridged, organometallic [2Fe-2S] ([2Fe]H) subsite. Profoundly different geometric models of the H-cluster redox states that orchestrate the electron/proton transfer steps of H2 bond activation have been proposed. We have examined this question in the [FeFe] H2ase I from Clostridium acetobutylicum (CaI) by Fourier-transform infrared (FTIR) spectroscopy with temperature annealing and H/D isotope exchange to identify the relevant redox states and define catalytic transitions. One-electron reduction of Hox led to formation of HredH+ ([4Fe-4S]H2+-FeI-FeI) and Hred' ([4Fe-4S]H1+-FeII-FeI), with both states characterized by low frequency μ-CO IR modes consistent with a fully bridged [2Fe]H. Similar μ-CO IR modes were also identified for HredH+ of the [FeFe] H2ase from Chlamydomonas reinhardtii (CrHydA1). The CaI proton-transfer variant C298S showed enrichment of an H/D isotope-sensitive μ-CO mode, a component of the hydride bound H-cluster IR signal, Hhyd. Equilibrating CaI with increasing amounts of NaDT, and probed at cryogenic temperatures, showed HredH+ was converted to Hhyd. Over an increasing temperature range from 10 to 260 K catalytic turnover led to loss of Hhyd and appearance of Hox, consistent with enzymatic turnovermore » and H2 formation. The results show for CaI that the μ-CO of [2Fe]H remains bridging for all of the 'Hred' states and that HredH+ is on pathway to Hhyd and H2 evolution in the catalytic mechanism. Here, this provides a blueprint for designing small molecule catalytic analogs« less

Authors:
 [1];  [1];  [1];  [2];  [2]; ORCiD logo [1]
  1. National Renewable Energy Lab. (NREL), Golden, CO (United States)
  2. Colorado School of Mines, Golden, CO (United States)
Publication Date:
Research Org.:
National Renewable Energy Laboratory (NREL), Golden, CO (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1440310
Report Number(s):
NREL/JA-2700-71170
Journal ID: ISSN 0002-7863
Grant/Contract Number:  
AC36-08GO28308
Resource Type:
Accepted Manuscript
Journal Name:
Journal of the American Chemical Society
Additional Journal Information:
Journal Volume: 140; Journal Issue: 24; Journal ID: ISSN 0002-7863
Publisher:
American Chemical Society (ACS)
Country of Publication:
United States
Language:
English
Subject:
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; [FeFe]-hydrogenases; catalysis; catalytic transitions

Citation Formats

Ratzloff, Michael W., Artz, Jacob H., Mulder, David W., Collins, Reuben T., Furtak, Thomas E., and King, Paul W. CO-Bridged H-Cluster Intermediates in the Catalytic Mechanism of [FeFe]-Hydrogenase CaI. United States: N. p., 2018. Web. doi:10.1021/jacs.8b03072.
Ratzloff, Michael W., Artz, Jacob H., Mulder, David W., Collins, Reuben T., Furtak, Thomas E., & King, Paul W. CO-Bridged H-Cluster Intermediates in the Catalytic Mechanism of [FeFe]-Hydrogenase CaI. United States. https://doi.org/10.1021/jacs.8b03072
Ratzloff, Michael W., Artz, Jacob H., Mulder, David W., Collins, Reuben T., Furtak, Thomas E., and King, Paul W. Wed . "CO-Bridged H-Cluster Intermediates in the Catalytic Mechanism of [FeFe]-Hydrogenase CaI". United States. https://doi.org/10.1021/jacs.8b03072. https://www.osti.gov/servlets/purl/1440310.
@article{osti_1440310,
title = {CO-Bridged H-Cluster Intermediates in the Catalytic Mechanism of [FeFe]-Hydrogenase CaI},
author = {Ratzloff, Michael W. and Artz, Jacob H. and Mulder, David W. and Collins, Reuben T. and Furtak, Thomas E. and King, Paul W.},
abstractNote = {The [FeFe]-hydrogenases ([FeFe] H2ases) catalyze reversible H2 activation at the H-cluster, which is composed of a [4Fe-4S]H subsite linked by a cysteine thiolate to a bridged, organometallic [2Fe-2S] ([2Fe]H) subsite. Profoundly different geometric models of the H-cluster redox states that orchestrate the electron/proton transfer steps of H2 bond activation have been proposed. We have examined this question in the [FeFe] H2ase I from Clostridium acetobutylicum (CaI) by Fourier-transform infrared (FTIR) spectroscopy with temperature annealing and H/D isotope exchange to identify the relevant redox states and define catalytic transitions. One-electron reduction of Hox led to formation of HredH+ ([4Fe-4S]H2+-FeI-FeI) and Hred' ([4Fe-4S]H1+-FeII-FeI), with both states characterized by low frequency μ-CO IR modes consistent with a fully bridged [2Fe]H. Similar μ-CO IR modes were also identified for HredH+ of the [FeFe] H2ase from Chlamydomonas reinhardtii (CrHydA1). The CaI proton-transfer variant C298S showed enrichment of an H/D isotope-sensitive μ-CO mode, a component of the hydride bound H-cluster IR signal, Hhyd. Equilibrating CaI with increasing amounts of NaDT, and probed at cryogenic temperatures, showed HredH+ was converted to Hhyd. Over an increasing temperature range from 10 to 260 K catalytic turnover led to loss of Hhyd and appearance of Hox, consistent with enzymatic turnover and H2 formation. The results show for CaI that the μ-CO of [2Fe]H remains bridging for all of the 'Hred' states and that HredH+ is on pathway to Hhyd and H2 evolution in the catalytic mechanism. Here, this provides a blueprint for designing small molecule catalytic analogs},
doi = {10.1021/jacs.8b03072},
journal = {Journal of the American Chemical Society},
number = 24,
volume = 140,
place = {United States},
year = {2018},
month = {5}
}

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Figures / Tables:

Figure 1 Figure 1: Top. Structural representation of Clostridium acetobu-tylicum [FeFe] H2ase I (CaI) showing the convergence of electron (e-) and proton (H+) transfer pathways. The F-cluster (green) and H-cluster (blue) domains are based on the crystal structure of Clostridium pasteurianum [FeFe] H2ase I (CpI), PDB 3C8Y.24 Bottom. Models of the H-clustermore » HredH+ state. A. The [2Fe]H µ-CO shifted to a terminal position, with an open coordination site at Fed.16, 25 B. The [2Fe]H retains a µ-CO, with an open coordination site at Fed.7, 13, 22, 26 C. The [2Fe]H with a µ-H-, and a CO occupying the Fed open coordination site.27 For each model, one-electron reduction of HredH+ maintains the same [2Fe]H geometry, but [4Fe-4S]H oxidation state changes from 2+ to 1+ (note: the nomenclature and amine protonation state of [2Fe]H also varies in each model).16, 22, 27-28« less

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