HydF as a scaffold protein in [FeFe] hydrogenase H-cluster biosynthesis

McGlynn, Shawn E.; Shepard, Eric M.; Winslow, Mark A.; Naumov, Anatoli V.; Duschene, Kaitlin S.; Posewitz, Matthew C.; Broderick, William E.; Broderick, Joan B.; Peters, John W.

doi:10.1016/j.febslet.2008.04.063

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Volume 582, Issue 15 , Pages 2183-2187, 25 June 2008

HydF as a scaffold protein in [FeFe] hydrogenase H-cluster biosynthesis

Edited by Stuart Ferguson

Shawn E. McGlynn
- Department of Chemistry and Biochemistry, The Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT 59717, United States
Eric M. Shepard
- Department of Chemistry and Biochemistry, The Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT 59717, United States
Mark A. Winslow
- Department of Chemistry and Biochemistry, The Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT 59717, United States
Anatoli V. Naumov
- Department of Chemistry and Biochemistry, The Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT 59717, United States
Kaitlin S. Duschene
- Department of Chemistry and Biochemistry, The Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT 59717, United States
Matthew C. Posewitz
- Colorado School of Mines, Department of Chemistry and Geochemistry, Golden, CO 80401, United States
- National Renewable Energy Laboratory, Golden, CO 80401, United States
William E. Broderick
- Department of Chemistry and Biochemistry, The Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT 59717, United States
Joan B. Broderick
- Department of Chemistry and Biochemistry, The Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT 59717, United States
- Corresponding authors. Fax: +1 406 994 7470.
John W. Peters
- Department of Chemistry and Biochemistry, The Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT 59717, United States
- Corresponding authors. Fax: +1 406 994 7470.

Received 23 April 2008; accepted 24 April 2008. published online 22 May 2008.

Abstract

In an effort to determine the specific protein component(s) responsible for in vitro activation of the [FeFe] hydrogenase (HydA), the individual maturation proteins HydE, HydF, and HydG from Clostridium acetobutylicum were purified from heterologous expressions in Escherichia coli. Our results demonstrate that HydF isolated from a strain expressing all three maturation proteins is sufficient to confer hydrogenase activity to purified inactive heterologously expressed HydA (expressed in the absence of HydE, HydF, and HydG). These results represent the first in vitro maturation of [FeFe] hydrogenase with purified proteins, and suggest that HydF functions as a scaffold upon which an H-cluster intermediate is synthesized.

Abbreviations: HydA^ΔEFG, HydA expressed in the absence of HydE, HydF, and HydG, HydF^EG, HydF co-expressed with HydE and HydG, HydF^ΔEG, HydF expressed in the absence of HydE and HydG, AdoMet, S-adenosylmethionine