The maturase HydF enables [FeFe] hydrogenase assembly via transient, cofactor-dependent interactionsShow others and affiliations
2020 (English)In: Journal of Biological Chemistry, ISSN 0021-9258, E-ISSN 1083-351X, Vol. 295, no 33, p. 11891-11901Article in journal (Refereed) Published
Abstract [en]
[FeFe] hydrogenases have attracted extensive attention in the field of renewable energy research because of their remarkable efficiency for H(2)gas production. H(2)formation is catalyzed by a biologically unique hexanuclear iron cofactor denoted the H-cluster. The assembly of this cofactor requires a dedicated maturation machinery including HydF, a multidomain [4Fe4S] cluster protein with GTPase activity. HydF is responsible for harboring and delivering a precatalyst to the apo-hydrogenase, but the details of this process are not well understood. Here, we utilize gas-phase electrophoretic macromolecule analysis to show that a HydF dimer forms a transient interaction complex with the hydrogenase and that the formation of this complex depends on the cofactor content on HydF. Moreover, Fourier transform infrared, electron paramagnetic resonance, and UV-visible spectroscopy studies of mutants of HydF show that the isolated iron-sulfur cluster domain retains the capacity for binding the precatalyst in a reversible fashion and is capable of activating apo-hydrogenase inin vitroassays. These results demonstrate the central role of the iron-sulfur cluster domain of HydF in the final stages of H-cluster assembly,i.e.in binding and delivering the precatalyst.
Place, publisher, year, edition, pages
American Society for Biochemistry and Molecular Biology , 2020. Vol. 295, no 33, p. 11891-11901
Keywords [en]
metalloenzyme, metal ion-protein interaction, mass spectrometry (MS), Fourier transform IR (FTIR), hydrogenase, cofactor, chaperone, scaffold, iron-sulfur protein, protein-protein interaction, metallo-cofactor assembly
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-175107DOI: 10.1074/jbc.RA119.011419ISI: 000563922500041PubMedID: 32620553Scopus ID: 2-s2.0-85089787702OAI: oai:DiVA.org:umu-175107DiVA, id: diva2:1471721
2020-09-292020-09-292023-03-24Bibliographically approved