Cell-free H-cluster Synthesis and [FeFe] Hydrogenase Activation: All Five CO and CN– Ligands Derive from Tyrosine
Abstract
[FeFe] hydrogenases are promising catalysts for producing hydrogen as a sustainable fuel and chemical feedstock, and they also serve as paradigms for biomimetic hydrogen-evolving compounds. Hydrogen formation is catalyzed by the H-cluster, a unique iron-based cofactor requiring three carbon monoxide (CO) and two cyanide (CN–) ligands as well as a dithiolate bridge. Three accessory proteins (HydE, HydF, and HydG) are presumably responsible for assembling and installing the Hcluster, yet their precise roles and the biosynthetic pathway have yet to be fully defined. In this report, we describe effective cell-free methods for investigating H-cluster synthesis and [FeFe] hydrogenase activation. Combining isotopic labeling with FTIR spectroscopy, we conclusively show that each of the CO and CN– ligands derive respectively from the carboxylate and amino substituents of tyrosine. Such in vitro systems with reconstituted pathways comprise a versatile approach for studying biosynthetic mechanisms, and this work marks a significant step towards an understanding of both the proteinprotein interactions and complex reactions required for H-cluster assembly and hydrogenase maturation.
- Authors:
-
- Stanford University, CA (United States)
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States); University of California, Davis, CA (United States)
- University of California, Davis, CA (United States)
- Stanford University, CA (United States); Stanford University, CA (United States)
- Publication Date:
- Research Org.:
- Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER); National Institutes of Health (NIH); National Science Foundation (NSF)
- OSTI Identifier:
- 1627462
- Grant/Contract Number:
- AC02-05CH11231; GM-65440; CHE-0745353
- Resource Type:
- Accepted Manuscript
- Journal Name:
- PLoS ONE
- Additional Journal Information:
- Journal Volume: 6; Journal Issue: 5; Journal ID: ISSN 1932-6203
- Publisher:
- Public Library of Science
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; tyrosine; vibration; chemical synthesis; Fourier transform infrared spectroscopy; biochemical cofactors; chemical radicals; cysteine; catalysts
Citation Formats
Kuchenreuther, Jon M., George, Simon J., Grady-Smith, Celestine S., Cramer, Stephen P., and Swartz, James R. Cell-free H-cluster Synthesis and [FeFe] Hydrogenase Activation: All Five CO and CN– Ligands Derive from Tyrosine. United States: N. p., 2011.
Web. doi:10.1371/journal.pone.0020346.
Kuchenreuther, Jon M., George, Simon J., Grady-Smith, Celestine S., Cramer, Stephen P., & Swartz, James R. Cell-free H-cluster Synthesis and [FeFe] Hydrogenase Activation: All Five CO and CN– Ligands Derive from Tyrosine. United States. https://doi.org/10.1371/journal.pone.0020346
Kuchenreuther, Jon M., George, Simon J., Grady-Smith, Celestine S., Cramer, Stephen P., and Swartz, James R. Tue .
"Cell-free H-cluster Synthesis and [FeFe] Hydrogenase Activation: All Five CO and CN– Ligands Derive from Tyrosine". United States. https://doi.org/10.1371/journal.pone.0020346. https://www.osti.gov/servlets/purl/1627462.
@article{osti_1627462,
title = {Cell-free H-cluster Synthesis and [FeFe] Hydrogenase Activation: All Five CO and CN– Ligands Derive from Tyrosine},
author = {Kuchenreuther, Jon M. and George, Simon J. and Grady-Smith, Celestine S. and Cramer, Stephen P. and Swartz, James R.},
abstractNote = {[FeFe] hydrogenases are promising catalysts for producing hydrogen as a sustainable fuel and chemical feedstock, and they also serve as paradigms for biomimetic hydrogen-evolving compounds. Hydrogen formation is catalyzed by the H-cluster, a unique iron-based cofactor requiring three carbon monoxide (CO) and two cyanide (CN–) ligands as well as a dithiolate bridge. Three accessory proteins (HydE, HydF, and HydG) are presumably responsible for assembling and installing the Hcluster, yet their precise roles and the biosynthetic pathway have yet to be fully defined. In this report, we describe effective cell-free methods for investigating H-cluster synthesis and [FeFe] hydrogenase activation. Combining isotopic labeling with FTIR spectroscopy, we conclusively show that each of the CO and CN– ligands derive respectively from the carboxylate and amino substituents of tyrosine. Such in vitro systems with reconstituted pathways comprise a versatile approach for studying biosynthetic mechanisms, and this work marks a significant step towards an understanding of both the proteinprotein interactions and complex reactions required for H-cluster assembly and hydrogenase maturation.},
doi = {10.1371/journal.pone.0020346},
journal = {PLoS ONE},
number = 5,
volume = 6,
place = {United States},
year = {Tue May 31 00:00:00 EDT 2011},
month = {Tue May 31 00:00:00 EDT 2011}
}
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