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His-Ligation to the [4Fe–4S] Subcluster Tunes the Catalytic Bias of [FeFe] Hydrogenase

Rodríguez-Maciá, Patricia, Kertess, Leonie, Burnik, Jan, Birrell, James A., Hofmann, Eckhard, Lubitz, Wolfgang, Happe, Thomas, Rüdiger, Olaf
Journal of the American Chemical Society 2018 v.141 no.1 pp. 472-481
Chlamydomonas reinhardtii, active sites, chemical bonding, cysteine, electron transfer, electrons, energy, enzymes, histidine, hydrogen, hydrogen production, injection site, oxidation, protons, redox potential, thermodynamics
[FeFe] hydrogenases interconvert H₂ into protons and electrons reversibly and efficiently. The active site H-cluster is composed of two sites: a unique [2Fe] subcluster ([2Fe]H) covalently linked via cysteine to a canonical [4Fe–4S] cluster ([4Fe–4S]H). Both sites are redox active and electron transfer is proton-coupled, such that the potential of the H-cluster lies very close to the H₂ thermodynamic potential, which confers the enzyme with the ability to operate quickly in both directions without energy losses. Here, one of the cysteines coordinating [4Fe–4S]H (Cys362) in the [FeFe] hydrogenase from the green algae Chlamydomonas reinhardtii (CrHydA1) was exchanged with histidine and the resulting C362H variant was shown to contain a [4Fe–4S] cluster with a more positive redox potential than the wild-type. The change in the [4Fe–4S] cluster potential resulted in a shift of the catalytic bias, diminishing the H₂ production activity but giving significantly higher H₂ oxidation activity, albeit with a 200 mV overpotential requirement. These results highlight the importance of the [4Fe–4S] cluster as an electron injection site, modulating the redox potential and the catalytic properties of the H-cluster.