Hydrogenases in Desulfovibrio vulgaris Hildenborough: structural and physiologic characterisation of the membrane-bound [NiFeSe] hydrogenase

Valente, Filipa M. A.; Oliveira, A. Sofia F.; Gnadt, Nicole; Pacheco, Isabel; Coelho, Ana V.; Xavier, Ant�nio V.; Teixeira, Miguel; Soares, Cl�udio M.; Pereira, In�s A. C.

Abstract

The genome of Desulfovibrio vulgaris Hildenborough (DvH) encodes for six hydrogenases (Hases), making it an interesting organism to study the role of these proteins in sulphate respiration. In this work we address the role of the [NiFeSe] Hase, found to be the major Hase associated with the cytoplasmic membrane. The purified enzyme displays interesting catalytic properties, such as a very high H2 production activity, which is dependent on the presence of phospholipids or detergent, and resistance to oxygen inactivation since it is isolated aerobically in a Ni(II) oxidation state. Evidence was obtained that the [NiFeSe] Hase is post-translationally modified to include a hydrophobic group bound to the N-terminal, which is responsible for its membrane association. Cleavage of this group originates a soluble, less active form of the enzyme. Sequence analysis shows that [NiFeSe] Hases from Desulfovibrionacae form a separate family from the [NiFe] enzymes of these organisms, and are more closely related to [NiFe] Hases from more distant bacterial species that have a medial [4Fe4S]2+/1+ cluster, but not a selenocysteine. The interaction of the [NiFeSe] Hase with periplasmic cytochromes was investigated and is similar to the [NiFe]1 Hase, with the Type I cytochrome c3 as the preferred electron acceptor. A model of the DvH [NiFeSe] Hase was generated based on the structure of the Desulfomicrobium baculatum enzyme. The structures of the two [NiFeSe] Hases are compared with the structures of [NiFe] Hases, to evaluate the consensual structural differences between the two families. Several conserved residues close to the redox centres were identified, which may be relevant to the higher activity displayed by [NiFeSe] Hases.

Citation

Valente, F. M. A., Oliveira, A. S. F., Gnadt, N., Pacheco, I., Coelho, A. V., Xavier, A. V., …Pereira, I. A. C. (2005). Hydrogenases in Desulfovibrio vulgaris Hildenborough: structural and physiologic characterisation of the membrane-bound [NiFeSe] hydrogenase. Journal of biological inorganic chemistry : JBIC : a publication of the Society of Biological Inorganic Chemistry, 10(6), 667-682. https://doi.org/10.1007/s00775-005-0022-4

• Journal Article Type: Article
• Acceptance Date: Oct 1, 2005
• Online Publication Date: Sep 27, 2005
• Publication Date: 2005-10
• Deposit Date: Aug 1, 2016
• Journal: Journal of Biological Inorganic Chemistry
• Electronic ISSN: 0949-8257
• Publisher: BMC
• Peer Reviewed: Peer Reviewed
• Volume: 10
• Issue: 6
• Pages: 667-682
• DOI: https://doi.org/10.1007/s00775-005-0022-4
• Keywords: Hydrogenases, Cytochromes, Sulphate-reducing bacteria, Electron transfer
• Public URL: http://researchrepository.napier.ac.uk/Output/321321