TY - JOUR
T1 - The direct role of selenocysteine in [NiFeSe] hydrogenase maturation and catalysis
AU - Marques, Marta C.
AU - Tapia, Cristina
AU - Gutiérrez-Sanz, Oscar
AU - Ramos, Ana Raquel
AU - Keller, Kimberly L.
AU - Wall, Judy D.
AU - De Lacey, Antonio L.
AU - Matias, Pedro M.
AU - Pereira, Inês A.C.
PY - 2017/5/1
Y1 - 2017/5/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85015672149&partnerID=8YFLogxK
U2 - 10.1038/nchembio.2335
DO - 10.1038/nchembio.2335
M3 - Article
C2 - 28319099
AN - SCOPUS:85015672149
VL - 13
SP - 544
EP - 550
JO - Nature Chemical Biology
JF - Nature Chemical Biology
SN - 1552-4450
IS - 5
ER -