title: HydF as a scaffold protein in [FeFe] hydrogenase H-cluster biosynthesis.
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.
authors.affiliation: Department of Chemistry and Biochemistry, The Astrobiology Biogeocatalysis Research Center, Montana State University, Bozeman, MT 59717, USA.
authors.forename: Shawn E
authors.initials: SE
authors.lastname: McGlynn
authors.forename: Eric M
authors.initials: EM
authors.lastname: Shepard
authors.forename: Mark A
authors.initials: MA
authors.lastname: Winslow
authors.forename: Anatoli V
authors.initials: AV
authors.lastname: Naumov
authors.forename: Kaitlin S
authors.initials: KS
authors.lastname: Duschene
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Posewitz
authors.forename: William E
authors.initials: WE
authors.lastname: Broderick
authors.forename: Joan B
authors.initials: JB
authors.lastname: Broderick
authors.forename: John W
authors.initials: JW
authors.lastname: Peters
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D003001:Cloning, Molecular; D046969:Clostridium acetobutylicum / Q000201:enzymology*; D004926:Escherichia coli / Q000235:genetics; D006864:Hydrogenase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D007506:Iron-Sulfur Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D007506:Iron-Sulfur Proteins; D011994:Recombinant Proteins; C118830:iron hydrogenase; D006864:Hydrogenase