1FRF

CRYSTAL STRUCTURE OF THE NI-FE HYDROGENASE FROM DESULFOVIBRIO FRUCTOSOVORANS

Summary for 1FRF

• Entry DOI: 10.2210/pdb1frf/pdb
• Descriptor: [NI-FE] HYDROGENASE, IRON/SULFUR CLUSTER, FE3-S4 CLUSTER, ... (8 entities in total)
• Functional Keywords: ni-fe hydrogenase, oxidoreductase
• Biological source: Desulfovibrio fructosovorans; More
• Cellular location: Periplasm: P18187 P18188
• Total number of polymer chains: 2
• Total formula weight: 91229.81

Authors

Montet, Y.,Volbeda, A.,Piras, C.,Hatchikian, E.C.,Frey, M.,Fontecilla, J.C. (deposition date: 1998-07-21, release date: 1998-07-29, Last modification date: 2023-08-09)

Primary citation

Rousset, M.,Montet, Y.,Guigliarelli, B.,Forget, N.,Asso, M.,Bertrand, P.,Fontecilla-Camps, J.C.,Hatchikian, E.C.
3Fe-4S] to [4Fe-4S] cluster conversion in Desulfovibrio fructosovorans [NiFe] hydrogenase by site-directed mutagenesis
Proc.Natl.Acad.Sci.USA, 95:11625-11630, 1998

PubMed Abstract: The role of the high potential [3Fe-4S]1+,0 cluster of [NiFe] hydrogenase from Desulfovibrio species located halfway between the proximal and distal low potential [4Fe-4S]2+,1+ clusters has been investigated by using site-directed mutagenesis. Proline 238 of Desulfovibrio fructosovorans [NiFe] hydrogenase, which occupies the position of a potential ligand of the lacking fourth Fe-site of the [3Fe-4S] cluster, was replaced by a cysteine residue. The properties of the mutant enzyme were investigated in terms of enzymatic activity, EPR, and redox properties of the iron-sulfur centers and crystallographic structure. We have shown on the basis of both spectroscopic and x-ray crystallographic studies that the [3Fe-4S] cluster of D. fructosovorans hydrogenase was converted into a [4Fe-4S] center in the P238 mutant. The [3Fe-4S] to [4Fe-4S] cluster conversion resulted in a lowering of approximately 300 mV of the midpoint potential of the modified cluster, whereas no significant alteration of the spectroscopic and redox properties of the two native [4Fe-4S] clusters and the NiFe center occurred. The significant decrease of the midpoint potential of the intermediate Fe-S cluster had only a slight effect on the catalytic activity of the P238C mutant as compared with the wild-type enzyme. The implications of the results for the role of the high-potential [3Fe-4S] cluster in the intramolecular electron transfer pathway are discussed.

PubMed: 9751716
DOI: 10.1073/pnas.95.20.11625

Experimental method

X-RAY DIFFRACTION (2.7 Å)