English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/189723
Share/Impact:
Statistics
logo share SHARE logo core CORE   Add this article to your Mendeley library MendeleyBASE

Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL
Exportar a otros formatos:

Title

A threonine stabilizes the NiC and NiR catalytic intermediates of [NiFe]-hydrogenase

AuthorsAbou-Hamdan, A.; Ceccaldi, P.; Lebrette, Hugo; Gutiérrez-Sanz, Óscar ; Richaud, P.; Cournac, L.; Guigliarelli, Bruno; López de Lacey, Antonio ; Léger, Christophe; Volbeda, A.; Burlat, Bénédicte; Dementin, Sébastien
Issue Date27-Mar-2015
PublisherAmerican Society for Biochemistry and Molecular Biology
CitationJournal of Biological Chemistry 290(13): 8550-8558 (2015)
AbstractThe heterodimeric [NiFe] hydrogenase from Desulfovibrio fructosovorans catalyzes the reversible oxidation of H2 into protons and electrons. The catalytic intermediates have been attributed to forms of the active site (NiSI, NiR, and NiC) detected using spectroscopic methods under potentiometric but non-catalytic conditions. Here, we produced variants by replacing the conserved Thr-18 residue in the small subunit with Ser, Val, Gln, Gly, or Asp, and we analyzed the effects of these mutations on the kinetic (H2 oxidation, H2 production, and H/D exchange), spectroscopic (IR, EPR), and structural properties of the enzyme. The mutations disrupt the H-bond network in the crystals and have a strong effect on H2 oxidation and H2 production turnover rates. However, the absence of correlation between activity and rate of H/D exchange in the series of variants suggests that the alcoholic group of Thr-18 is not necessarily a proton relay. Instead, the correlation between H2 oxidation and production activity and the detection of the NiC species in reduced samples confirms that NiC is a catalytic intermediate and suggests that Thr-18 is important to stabilize the local protein structure of the active site ensuring fast NiSI-NiC-NiR interconversions during H2 oxidation/production.
DescriptionEsta investigación se publicó originalmente en el Journal of Biological Chemistry. Abou-Hamdan, A.; Ceccaldi, P.; Lebrette, Hugo; Gutiérrez-Sanz, Óscar ; Richaud, P.; Cournac, L.; Guigliarelli, Bruno; López de Lacey, Antonio ; Léger, Christophe; Volbeda, A.; Burlat, Bénédicte; Dementin, Sébastien. A threonine stabilizes the NiC and NiR catalytic intermediates of [NiFe]-hydrogenase. J. Biol. Chem 2015; 290(13): 8550-8558. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.
Publisher version (URL)https://doi.org/10.1074/jbc.M114.630491
URIhttp://hdl.handle.net/10261/189723
DOI10.1074/jbc.M114.630491
ISSN0021-9258
E-ISSN1083-351X
Appears in Collections:(ICP) Artículos
Files in This Item:
File Description SizeFormat 
Threonine_Abou_Hamdan_2015.pdf1,83 MBAdobe PDFThumbnail
View/Open
Show full item record
Review this work
 

Related articles:


WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.