An in vitro reconstitution system to monitor iron transfer to the active site during the maturation of [NiFe]-hydrogenase.

ABSTRACT: [NiFe]-hydrogenases (Hyds) comprise a small and a large subunit. The latter harbors the biologically unique [NiFe](CN)₂CO active-site cofactor. The maturation process includes the assembly of the [Fe](CN)₂CO cofactor precursor, nickel binding, endoproteolytic cleavage of the large subunit, and dimerization with the small subunit to yield active enzyme. The biosynthesis of the [Fe](CN)₂CO moiety of [NiFe]-Hyd-1 and Hyd-2 occurs on the scaffold complex HybG-HypD (GD), whereas the HypC-HypD complex is specific for the assembly of Hyd-3. The metabolic source and the route for delivering iron to the active site remain unclear. To investigate the maturation process of O₂-tolerant Hyd-1 from Escherichia coli, we developed an enzymatic in vitro reconstit

SUBMITTER: Soboh B 

PROVIDER: S-EPMC9418501 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature