A structure-based site-directed mutagenesis study on the neurolysin (EC 3.4.24.16) and thimet oligopeptidase (EC 3.4.24.15) catalysis

A structure-based site-directed mutagenesis study on the neurolysin (EC 3.4.24.16) and thimet oligopeptidase (EC 3.4.24.15) catalysis

Author Oliveira, Vitor Autor UNIFESP Google Scholar
Araujo, M. C. Google Scholar
Rioli, V Google Scholar
Camargo, Antonio Carlos Martins de Autor UNIFESP Google Scholar
Tersariol, Ivarne Luis dos Santos Autor UNIFESP Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Ferro, Emer Suavinho Autor UNIFESP Google Scholar
Institution Univ Mogi das Cruzes
Inst Butantan
Universidade de São Paulo (USP)
Universidade Federal de São Paulo (UNIFESP)
Abstract Neurolysin (EP24.16) and thimet oligopeptidase (EP24.15) are closely related metalloendopeptidases. Site-directed mutagenesis of Tyr(613) (EP24.16) or Tyr(612) (EP24.15) to either Phe or Ala promoted a strong reduction of k(cat)/K-M for both enzymes. These data suggest the importance of both hydroxyl group and aromatic ring at this specific position during substrate hydrolysis by these peptidases. Furthermore, the EP24.15 A607G mutant showed a k(cat)/K-M of 2x10(5) M-1 s(-1) for the Abz-GFSIFRQ-EDDnp substrate, similar to that of EP24.16 (k(cat)/K-M = 3x10(5) M-1 s(-1)) which contains Gly at the corresponding position; the wild type EP24.15 has a k(cat)/K-M of 2.5x10(4) M-1 s(-1) for this substrate. (C) 2003 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
Keywords enzyme specificity
catalytic mechanism
site-directed mutagenesis
Language English
Date 2003-04-24
Published in Febs Letters. Amsterdam: Elsevier B.V., v. 541, n. 1-3, p. 89-92, 2003.
ISSN 0014-5793 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 89-92
Origin http://dx.doi.org/10.1016/S0014-5793(03)00310-7
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000182481600017
URI http://repositorio.unifesp.br/handle/11600/27211

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