Description:
<jats:title>Abstract</jats:title><jats:p>The structure of the radical <jats:italic>S</jats:italic>‐adenosyl‐<jats:sc>L</jats:sc>‐methionine (SAM) [FeFe]‐hydrogenase maturase HydG involved in CN<jats:sup>−</jats:sup>/CO synthesis is characterized by two internal tunnels connecting its tyrosine‐binding pocket with the external medium and the C‐terminal Fe<jats:sub>4</jats:sub>S<jats:sub>4</jats:sub> cluster‐containing region. A comparison with a tryptophan‐bound NosL structure suggests that substrate binding causes the closing of the first tunnel and, along with mutagenesis studies, that tyrosine binds to HydG with its amino group well positioned for H‐abstraction by SAM. In this orientation the dehydroglycine (DHG) fragment caused by tyrosine CαCβ bond scission can readily migrate through the second tunnel towards the C‐terminal domain where both CN<jats:sup>−</jats:sup> and CO are synthesized. Our HydG structure appears to be in a relaxed state with its C‐terminal cluster CysX<jats:sub>2</jats:sub>CysX<jats:sub>22</jats:sub>Cys motif exposed to solvent. A rotation of this domain coupled to Fe<jats:sub>4</jats:sub>S<jats:sub>4</jats:sub> cluster assembly would bury its putatively reactive unique Fe ion thereby allowing it to interact with DHG.</jats:p>