Semisynthetic maturation of [FeFe]-hydrogenase using [Fe 2 (μ-SH) 2 (CN) 2 (CO) 4 ] 2- : key roles for HydF and GTP.

Here we describe maturation of the [FeFe]-hydrogenase from its [4Fe-4S]-bound precursor state by using the synthetic complex [Fe 2 (μ-SH) 2 (CN) 2 (CO) 4 ] 2- together with HydF and components of the glycine cleavage system, but in the absence of the maturases HydE and HydG. This semisynthetic and fully-defined maturation provides new insights into the nature of H-cluster biosynthesis.