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

• Autor(a) principal: Oliveira, Vitor [UNIFESP]
• Data de Publicação: 2003
• Outros Autores: Araujo, M. C., Rioli, V, Camargo, Antonio Carlos Martins de [UNIFESP], Tersariol, Ivarne Luis dos Santos [UNIFESP], Juliano, Maria Aparecida [UNIFESP], Juliano, Luiz [UNIFESP], Ferro, Emer Suavinho [UNIFESP]
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UNIFESP
• Texto Completo: http://dx.doi.org/10.1016/S0014-5793(03)00310-7; http://repositorio.unifesp.br/handle/11600/27211
• topic: enzyme specificity; catalytic mechanism; site-directed mutagenesis

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.