O2‑Tolerant H2 Activation by an Isolated Large Subunit of a [NiFe] Hydrogenase

• Published in: Biochemistry (Easton) 2018-09, Vol.57 (36), p.5339-5349
• Main Authors: Hartmann, Sven, Frielingsdorf, Stefan, Ciaccafava, Alexandre, Lorent, Christian, Fritsch, Johannes, Siebert, Elisabeth, Priebe, Jacqueline, Haumann, Michael, Zebger, Ingo, Lenz, Oliver
• Format: Article
• Language: English
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/acs.biochem.8b00760

The catalytic properties of hydrogenases are nature’s answer to the seemingly simple reaction H2 ⇌ 2H+ + 2e–. Members of the phylogenetically diverse subgroup of [NiFe] hydrogenases generally consist of at least two subunits, where the large subunit harbors the H2-activating [NiFe] site and the small subunit contains iron–sulfur clusters mediating e– transfer. Typically, [NiFe] hydrogenases are susceptible to inhibition by O2. Here, we conducted system minimization by isolating and analyzing the large subunit of one of the rare members of the group of O2-tolerant [NiFe] hydrogenases, namely the preHoxG protein of the membrane-bound hydrogenase from Ralstonia eutropha. Unlike previous assumptions, preHoxG was able to activate H2 as it clearly performed catalytic hydrogen/deuterium exchange. However, it did not execute the entire catalytic cycle described for [NiFe] hydrogenases. Remarkably, H2 activation was performed by preHoxG even in the presence of O2, although the unique [4Fe-3S] cluster located in the small subunit and described to be crucial for tolerance toward O2 was absent. These findings challenge the current understanding of O2 tolerance of [NiFe] hydrogenases. The applicability of this minimal hydrogenase in basic and applied research is discussed.