Site-directed mutagenesis and molecular modelling studies show the role of Asp82 and cysteines in rat acylase 1, a member of the M20 family(486 views) Herga S, Brutus A, Vitale RM, Miche H, Perrier J, Puigserver A, Scaloni A, Giardina T
Biochem Biophys Res Commun (ISSN: 0006-291xprint, 0006-291xlinking), 2005 May 6; 330(2): 540-546.
Paper type: Journal Article, Research Support, Non-U. S. Gov'T,
Impact factor: 3, 5-year impact factor: 2.474
Url: Not available.
Keywords: Amidohydrolases, Chemistry, Genetics, Metabolism, Animals, Aspartic Acid, Base Sequence, Cysteine, Dna Primers, Kidney, Enzymology, Models, Molecular, Mutagenesis, Site-Directed, Rats,
Affiliations: *** IBB - CNR ***
Institut Mediterraneen de Recherche en Nutrition, Laboratoire de Biochimie et Biologie de la Nutrition, UMR Universite Paul Cezanne Aix Marseille III-INRA 1111, service 342, Faculte des Sciences et Techniques Saint-Jerome, 13397 Marseille Cedex 20, France.,
References: Not available.
Site-directed mutagenesis and molecular modelling studies show the role of Asp82 and cysteines in rat acylase 1, a member of the M20 family
Acylase 1 from rat kidney catalyzes the hydrolysis of acyl-amino acids. Sequence alignment has shown that this enzyme belongs to the metalloprotein family M20. Site-directed mutagenesis experiments led to the identification of one functionally important amino acid residue located near one of the zinc coordinating residues, which play a critical role in the enzymatic activity. The D82N- and D82E-substituted forms showed no significant activity and very low activity, respectively, along with a loss of zinc coordination. Molecular modelling investigations indicated a putative role of D82 in ensuring a proper protonation of catalytic histidine. In addition, none of the five cysteine residues present in the rat kidney acylase 1 sequence seemed involved in the catalytic process: the loss of activity induced by the C294A substitution was probably due to a conformational change in the 3D structure.
Site-directed mutagenesis and molecular modelling studies show the role of Asp82 and cysteines in rat acylase 1, a member of the M20 family
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