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The direct role of selenocysteine in [NiFeSe] hydrogenase maturation and catalysis

DOI: 10.1038/nchembio.2335 DOI Help

Authors: Marta C. Marques (Universidade Nova de Lisboa) , Cristina Tapia (Instituto de Catálisis y Petroleoquímica (CSIC)) , Oscar Gutiérrez-sanz (Instituto de Catálisis y Petroleoquímica (CSIC)) , Ana Raquel Ramos (Universidade Nova de Lisboa) , Kimberly L. Keller (Ecosystems and Networks Integrated with Genes and Molecular Assemblies (ENIGMA)) , Judy D. Wall (University of Missouri) , Antonio L. De Lacey (Instituto de Catálisis y Petroleoquímica (CSIC)) , Pedro M. Matias (Institudo Tecnologia Quimica e Biologica, , Universidade Nova de Lisboa; Instituto de Biologia Experimental e Tecnológica (iBET)) , Inês A. C. Pereira (Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Chemical Biology , VOL 2011

State: Published (Approved)
Published: March 2017
Diamond Proposal Number(s): 10515

Abstract: Hydrogenases are highly active enzymes for hydrogen production and oxidation. [NiFeSe] hydrogenases, where selenocysteine is a ligand to the active site Ni, have high catalytic activities and a bias for H2 production. In contrast to [NiFe] hydrogenases, they display reduced H2 inhibition and are rapidly reactivated after contact with oxygen. Here, we report a homologous expression system for production of recombinant [NiFeSe] hydrogenase from Desulfovibrio vulgaris Hildenborough, and study of a Sec489Cys variant where for the first time a [NiFeSe] hydrogenase was converted to a [NiFe] type. This modification led to a severely reduced Ni incorporation, revealing the direct involvement of this residue in the maturation process. The Ni-depleted protein could be partly reconstituted to generate an enzyme displaying much reduced activity and inactive states characteristic of [NiFe] hydrogenases. The Ni-U489C variant shows that selenium plays a crucial role in protection against oxidative damage and the high catalytic activities of the [NiFeSe] hydrogenases.

Subject Areas: Biology and Bio-materials, Energy, Environment

Instruments: I02-Macromolecular Crystallography

Other Facilities: ESRF and SLS