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

Marta C. Marques, Cristina Tapia, Oscar Gutiérrez-Sanz, Ana Raquel Ramos, Kimberly L. Keller, Judy D. Wall, Antonio L. De Lacey, Pedro M. Matias, Inês A.C. Pereira

Research output: Contribution to journalArticlepeer-review

39 Citations (Scopus)

Abstract

Hydrogenases are highly active enzymes for hydrogen production and oxidation. [NiFeSe] hydrogenases, in which selenocysteine is a ligand to the active site Ni, have high catalytic activity 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 an expression system for production of recombinant [NiFeSe] hydrogenase from Desulfovibrio vulgaris Hildenborough and study of a selenocysteine-to-cysteine variant (Sec489Cys) in which, for the first time, a [NiFeSe] hydrogenase was converted to a [NiFe] type. This modification led to 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 showing much lower activity and inactive states characteristic of [NiFe] hydrogenases. The Ni-Sec489Cys variant shows that selenium has a crucial role in protection against oxidative damage and the high catalytic activities of the [NiFeSe] hydrogenases.

Original languageEnglish
Pages (from-to)544-550
Number of pages7
JournalNature Chemical Biology
Volume13
Issue number5
DOIs
Publication statusPublished - 1 May 2017

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