Differences in transition state stabilization between thermolysin (EC 3.4.24.27) and neprilysin (EC 3.4.24.11)

FEBS Letters

Marie-Claire, Cynthia¹  Tiraboschi, Gilles¹  Ruffet, Emmanuel¹  Fournie-Zaluski, Marie-Claude¹ 

[1] Département de Pharmacochimie Moléculaire et Structurale, U266 INSERM, URA D1500 CNRS, UFR des Sciences Pharmaceutiques et Biologiques, Faculté de Pharmacie, 4, Avenue de l'Observatoire, 75270 Paris Cedex 06, France

关键词: Neprilysin;    Thermolysin;    Site-directed mutagenesis;    Molecular modeling;    Phosphoramidon;    N-(α-rhamnopyranosyl-(oxyhydroxyphosphinyl))-l-leucyl-1-tryptophan;    thiorphan;    N-(2-(R;    S)-mercaptomethyl)-1-oxo-3-phenylpropyl)glycine;    ECE;    endothelin converting enzyme;    HACBO-Gly;    N-(2-(R;    S)-3-hydroxyaminocarbonyl-2-benzyl-1-oxopropyl)-glycine;   

DOI  :  10.1016/S0014-5793(98)01267-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Important homologies in the topology of the catalytic site and the mechanism of action of thermolysin and neprilysin have been evidenced by site-directed mutagenesis. The determination of differences in transition state stabilization between these peptidases could facilitate the design of specific inhibitors. Thus, two residues of thermolysin which could be directly (Tyr¹⁵⁷) or indirectly (Asp²²⁶) involved in the stabilization of the transition state and their putative counterparts in neprilysin (Tyr⁶²⁵ and Asp⁷⁰⁹) have been mutated. The results show that Tyr¹⁵⁷ is important for thermolysin activity while Tyr⁶²⁵ has no functional role in neprilysin. Conversely, the mutation of Asp²²⁶ induced a slight perturbation of thermolysin activity while Asp⁷⁰⁹ in neprilysin seems crucial in neprilysin catalysis. Taken together these data seem to indicate differences in the transition state mode of stabilization in the two peptidases.

【 授权许可】

Unknown