[NiFe] hydrogenases are complex model enzymes for the reversible cleavage of dihydrogen (H2 ). However, structural determinants of efficient H2 binding to their [NiFe] active site are not properly understood. Here, we present crystallographic and vibrational-spectroscopic insights into the unexplored structure of the H2 -binding [NiFe] intermediate. Using an F420 -reducing [NiFe]-hydrogenase from Methanosarcina barkeri as a model enzyme, we show that the protein backbone provides a strained chelating scaffold that tunes the [NiFe] active site for efficient H2 binding and conversion. The protein matrix also directs H2 diffusion to the [NiFe] site via two gas channels and allows the distribution of electrons between functional protomers through a subunit-bridging FeS cluster. Our findings emphasize the relevance of an atypical Ni coordination, thereby providing a blueprint for the design of bio-inspired H2 -conversion catalysts.
Keywords: [NiFe] hydrogenase; biocatalysis; crystal structure; hydrogen activation; vibrational spectroscopy.
© 2019 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.