New Insights into [FeFe] Hydrogenase Activation and Maturase Function

Kuchenreuther, Jon M.; Britt, R. David; Swartz, James R.

doi:10.1371/journal.pone.0045850

New Insights into [FeFe] Hydrogenase Activation and Maturase Function

Journal Article · Tue Sep 25 00:00:00 EDT 2012 · PLoS ONE

DOI:https://doi.org/10.1371/journal.pone.0045850· OSTI ID:1904544

Kuchenreuther, Jon M. ^[1]; Britt, R. David ^[2]; Swartz, James R. ^[3]

Stanford Univ., CA (United States); Univ. of California, Davis, CA (United States); Stanford Univ., CA
Univ. of California, Davis, CA (United States)
Stanford Univ., CA (United States)

[FeFe] hydrogenases catalyze H₂ production using the H-cluster, an iron-sulfur cofactor that contains carbon monoxide (CO), cyanide (CN^–), and a dithiolate bridging ligand. The HydE, HydF, and HydG maturases assist in assembling the H-cluster and maturing hydrogenases into their catalytically active form. Characterization of these maturases and in vitro hydrogenase activation methods have helped elucidate steps in the H-cluster biosynthetic pathway such as the HydG-catalyzed generation of the CO and CN^– ligands from free tyrosine. We have refined our cell-free approach for H-cluster synthesis and hydrogenase maturation by using separately expressed and purified HydE, HydF, and HydG. In this report, we illustrate how substrates and protein constituents influence hydrogenase activation, and for the first time, we show that each maturase can function catalytically during the maturation process. With precise control over the biomolecular components, we also provide evidence for H-cluster synthesis in the absence of either HydE or HydF, and we further show that hydrogenase activation can occur without exogenous tyrosine. Given these findings, we suggest a new reaction sequence for the [FeFe] hydrogenase maturation pathway. In our model, HydG independently synthesizes an iron-based compound with CO and CN^– ligands that is a precursor to the H-cluster [2Fe]_H subunit, and which we have termed HydG-co. We further propose that HydF is a transferase that stabilizes HydG-co and also shuttles the complete [2Fe]_H subcluster to the hydrogenase, a translocation process that may be catalyzed by HydE. In summary, this report describes the first example of reconstructing the [FeFe] hydrogenase maturation pathway using purified maturases and subsequently utilizing this in vitro system to better understand the roles of HydE, HydF, and HydG.

View Accepted Manuscript (DOE)

Research Organization:: Stanford Univ., CA (United States); Univ. of California, Davis, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Materials Sciences & Engineering Division (MSE)

Grant/Contract Number:: SC0002010; SC0004334

OSTI ID:: 1904544

Journal Information:: PLoS ONE, Journal Name: PLoS ONE Journal Issue: 9 Vol. 7; ISSN 1932-6203

Publisher:: Public Library of ScienceCopyright Statement

Country of Publication:: United States

Language:: English

References (31)

[FeFe]-Hydrogenase Cyanide Ligands Derived From S-Adenosylmethionine-Dependent Cleavage of Tyrosine Driesener, Rebecca C.; Challand, Martin R.; McGlynn, Shawn E. Angewandte Chemie International Edition, Vol. 49, Issue 9 https://doi.org/10.1002/anie.200907047	journal	January 2010
Further Characterization of the [FeFe]-Hydrogenase Maturase HydG Tron, Cécile; Cherrier, Mickaël V.; Amara, Patricia European Journal of Inorganic Chemistry, Vol. 2011, Issue 7 https://doi.org/10.1002/ejic.201001101	journal	February 2011
Biochemical characterization of the HydE and HydG iron-only hydrogenase maturation enzymes from Thermatoga maritima Rubach, Jon K.; Brazzolotto, Xavier; Gaillard, Jacques FEBS Letters, Vol. 579, Issue 22 https://doi.org/10.1016/j.febslet.2005.07.092	journal	August 2005
HydF as a scaffold protein in [FeFe] hydrogenase H-cluster biosynthesis McGlynn, Shawn E.; Shepard, Eric M.; Winslow, Mark A. FEBS Letters, Vol. 582, Issue 15 https://doi.org/10.1016/j.febslet.2008.04.063	journal	May 2008
The role of the maturase HydG in [FeFe]‐hydrogenase active site synthesis and assembly Pilet, Eric; Nicolet, Yvain; Mathevon, Carole FEBS Letters, Vol. 583, Issue 3 https://doi.org/10.1016/j.febslet.2009.01.004	journal	January 2009
The [FeFe]-hydrogenase maturase HydF from Clostridium acetobutylicum contains a CO and CN ⁻ ligated iron cofactor Czech, Ilka; Silakov, Alexey; Lubitz, Wolfgang FEBS Letters, Vol. 584, Issue 3 https://doi.org/10.1016/j.febslet.2009.12.016	journal	December 2009
A glycyl free radical as the precursor in the synthesis of carbon monoxide and cyanide by the [FeFe]-hydrogenase maturase HydG Nicolet, Yvain; Martin, Lydie; Tron, Cécile FEBS Letters, Vol. 584, Issue 19 https://doi.org/10.1016/j.febslet.2010.09.008	journal	September 2010
Desulfovibrio desulfuricans iron hydrogenase: the structure shows unusual coordination to an active site Fe binuclear center Nicolet, Yvain; Piras, Claudine; Legrand, Pierre Structure, Vol. 7, Issue 1, p. 13-23 https://doi.org/10.1016/s0969-2126(99)80005-7	journal	January 1999
Identification of Lysine 346 as a Functionally Important Residue for Pyridoxal 5‘-Phosphate Binding and Catalysis in Lysine 2,3-Aminomutase from Bacillus subtilis ^† Chen, Dawei; Frey, Perry A. Biochemistry, Vol. 40, Issue 2 https://doi.org/10.1021/bi002265w	journal	January 2001
Mechanism of Cysteine Desulfurase Slr0387 from Synechocystis sp. PCC 6803: Kinetic Analysis of Cleavage of the Persulfide Intermediate by Chemical Reductants Behshad, Elham; Parkin, Sara E.; Bollinger, J. Martin Biochemistry, Vol. 43, Issue 38 https://doi.org/10.1021/bi049143e	journal	September 2004
HscA and HscB Stimulate [2Fe-2S] Cluster Transfer from IscU to Apoferredoxin in an ATP-Dependent Reaction ^† Chandramouli, Kala; Johnson, Michael K. Biochemistry, Vol. 45, Issue 37 https://doi.org/10.1021/bi061237w	journal	September 2006
Formation and Properties of [4Fe-4S] Clusters on the IscU Scaffold Protein ^† Chandramouli, Kala; Unciuleac, Mihaela-Carmen; Naik, Sunil Biochemistry, Vol. 46, Issue 23 https://doi.org/10.1021/bi6026659	journal	June 2007
Pyrroloquinoline Quinone Biogenesis: Demonstration That PqqE from Klebsiella pneumoniae Is a Radical S -Adenosyl- l -methionine Enzyme Wecksler, Stephen R.; Stoll, Stefan; Tran, Ha Biochemistry, Vol. 48, Issue 42 https://doi.org/10.1021/bi900918b	journal	October 2009
[FeFe]-Hydrogenase Maturation: HydG-Catalyzed Synthesis of Carbon Monoxide Shepard, Eric M.; Duffus, Benjamin R.; George, Simon J. Journal of the American Chemical Society, Vol. 132, Issue 27 https://doi.org/10.1021/ja1012273	journal	July 2010
Dithiomethylether as a Ligand in the Hydrogenase H-Cluster Pandey, Arti S.; Harris, Travis V.; Giles, Logan J. Journal of the American Chemical Society, Vol. 130, Issue 13 https://doi.org/10.1021/ja711187e	journal	April 2008
Quantum Refinement of [FeFe] Hydrogenase Indicates a Dithiomethylamine Ligand Ryde, Ulf; Greco, Claudio; De Gioia, Luca Journal of the American Chemical Society, Vol. 132, Issue 13 https://doi.org/10.1021/ja909194f	journal	April 2010
Stepwise [FeFe]-hydrogenase H-cluster assembly revealed in the structure of HydAΔEFG Mulder, David W.; Boyd, Eric S.; Sarma, Ranjana Nature, Vol. 465, Issue 7295 https://doi.org/10.1038/nature08993	journal	April 2010
NifB-dependent in vitro synthesis of the iron-molybdenum cofactor of nitrogenase Curatti, L.; Ludden, P. W.; Rubio, L. M. Proceedings of the National Academy of Sciences, Vol. 103, Issue 14 https://doi.org/10.1073/pnas.0601115103	journal	March 2006
In vitro synthesis of the iron molybdenum cofactor of nitrogenase from iron, sulfur, molybdenum, and homocitrate using purified proteins Curatti, L.; Hernandez, J. A.; Igarashi, R. Y. Proceedings of the National Academy of Sciences, Vol. 104, Issue 45 https://doi.org/10.1073/pnas.0703050104	journal	October 2007
Synthesis of the 2Fe subcluster of the [FeFe]-hydrogenase H cluster on the HydF scaffold Shepard, E. M.; McGlynn, S. E.; Bueling, A. L. Proceedings of the National Academy of Sciences, Vol. 107, Issue 23 https://doi.org/10.1073/pnas.1001937107	journal	May 2010
Spectroscopic studies of ferricyanide oxidation of Azotobacter vinelandii ferredoxin I. Morgan, T. V.; Stephens, P. J.; Devlin, F. Proceedings of the National Academy of Sciences, Vol. 81, Issue 7 https://doi.org/10.1073/pnas.81.7.1931	journal	April 1984
The Requirement of Reductant for in Vitro Biosynthesis of the Iron-Molybdenum Cofactor of Nitrogenase Allen, Ronda M.; Chatterjee, Ranjini; Ludden, Paul W. Journal of Biological Chemistry, Vol. 271, Issue 8 https://doi.org/10.1074/jbc.271.8.4256	journal	February 1996
Discovery of Two Novel Radical S -Adenosylmethionine Proteins Required for the Assembly of an Active [Fe] Hydrogenase Posewitz, Matthew C.; King, Paul W.; Smolinski, Sharon L. Journal of Biological Chemistry, Vol. 279, Issue 24 https://doi.org/10.1074/jbc.m403206200	journal	April 2004
The [Fe-Fe]-Hydrogenase Maturation Protein HydF from Thermotoga maritima Is a GTPase with an Iron-Sulfur Cluster Brazzolotto, Xavier; Rubach, Jon K.; Gaillard, Jacques Journal of Biological Chemistry, Vol. 281, Issue 2 https://doi.org/10.1074/jbc.m510310200	journal	November 2005
Interplay of IscA and IscU in Biogenesis of Iron-Sulfur Clusters Yang, Juanjuan; Bitoun, Jacob P.; Ding, Huangen Journal of Biological Chemistry, Vol. 281, Issue 38 https://doi.org/10.1074/jbc.m601356200	journal	September 2006
Lysine 2,3‐aminomutase: is adenosylmethionine a poor man's adenosylcobalamin? Frey, Perry A. The FASEB Journal, Vol. 7, Issue 8 https://doi.org/10.1096/fasebj.7.8.8500691	journal	May 1993
Similarities in the Architecture of the Active Sites of Ni-Hydrogenases and Fe-Hydrogenases Detected by Means of Infrared Spectroscopy Spek, Trienke M.; Arendsen, Alexander F.; Happe, Randolph P. European Journal of Biochemistry, Vol. 237, Issue 3 https://doi.org/10.1111/j.1432-1033.1996.0629p.x	journal	May 1996
Pyridoxal Phosphate Enzymes: Mechanistic, Structural, and Evolutionary Considerations Eliot, Andrew C.; Kirsch, Jack F. Annual Review of Biochemistry, Vol. 73, Issue 1 https://doi.org/10.1146/annurev.biochem.73.011303.074021	journal	June 2004
Tyrosine, Cysteine, and S-Adenosyl Methionine Stimulate In Vitro [FeFe] Hydrogenase Activation Kuchenreuther, Jon M.; Stapleton, James A.; Swartz, James R. PLoS ONE, Vol. 4, Issue 10 https://doi.org/10.1371/journal.pone.0007565	journal	October 2009
High-Yield Expression of Heterologous [FeFe] Hydrogenases in Escherichia coli Kuchenreuther, Jon M.; Grady-Smith, Celestine S.; Bingham, Alyssa S. PLoS ONE, Vol. 5, Issue 11 https://doi.org/10.1371/journal.pone.0015491	journal	November 2010
Cell-free H-cluster Synthesis and [FeFe] Hydrogenase Activation: All Five CO and CN− Ligands Derive from Tyrosine Kuchenreuther, Jon M.; George, Simon J.; Grady-Smith, Celestine S. PLoS ONE, Vol. 6, Issue 5 https://doi.org/10.1371/journal.pone.0020346	journal	May 2011

Cited By (4)

[FeFe]- and [NiFe]-hydrogenase diversity, mechanism, and maturation Peters, John W.; Schut, Gerrit J.; Boyd, Eric S. Biochimica et Biophysica Acta (BBA) - Molecular Cell Research, Vol. 1853, Issue 6 https://doi.org/10.1016/j.bbamcr.2014.11.021	journal	June 2015
Electron Spin Relaxation and Biochemical Characterization of the Hydrogenase Maturase HydF: Insights into [2Fe-2S] and [4Fe-4S] Cluster Communication and Hydrogenase Activation Shepard, Eric M.; Byer, Amanda S.; Aggarwal, Priyanka Biochemistry, Vol. 56, Issue 25 https://doi.org/10.1021/acs.biochem.7b00169	journal	May 2017
Biochemical and Kinetic Characterization of Radical S -Adenosyl- l -methionine Enzyme HydG Driesener, Rebecca C.; Duffus, Benjamin R.; Shepard, Eric M. Biochemistry, Vol. 52, Issue 48 https://doi.org/10.1021/bi401143s	journal	November 2013
A [4Fe–4S]-Fe(CO)(CN)-l-cysteine intermediate is the first organometallic precursor in [FeFe] hydrogenase H-cluster bioassembly Rao, Guodong; Tao, Lizhi; Suess, Daniel L. M. Nature Chemistry, Vol. 10, Issue 5 https://doi.org/10.1038/s41557-018-0026-7	journal	April 2018

Similar Records

The [4Fe-4S] Cluster of HydF Is Essential for [FeFe]-Hydrogenase Maturation

Journal Article · Tue Dec 09 19:00:00 EST 2025 · Journal of the American Chemical Society · OSTI ID:3010489

H-cluster assembly intermediates built on HydF by the radical SAM enzymes HydE and HydG

Journal Article · Thu Sep 05 20:00:00 EDT 2019 · JBIC Journal of Biological Inorganic Chemistry · OSTI ID:1566047

[FeFe]-Hydrogenase: Defined Lysate-Free Maturation Reveals a Key Role for Lipoyl-H-Protein in DTMA Ligand Biosynthesis

Journal Article · Tue Mar 22 20:00:00 EDT 2022 · Angewandte Chemie (International Edition) · OSTI ID:1976282

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
biochemical cofactors
chemical radicals
chemical synthesis
cysteine
electron spin resonance spectroscopy
enzymes
small molecules
tyrosine

New Insights into [FeFe] Hydrogenase Activation and Maturase Function

Citation Formats

References (31)

Cited By (4)

Similar Records

Related Subjects