Investigation of neutral endopeptidases (EC 3.4.24.11) and of neutral proteinases (EC 3.4.24.4) using a new sensitive two‐stage enzymatic reaction

FEBS Letters

Indig, Fred E.¹  Ben-Meir, Daniella¹  Blumberg, Shmaryahu¹  Spungin, Anya¹ 

[1] Sackler Institute of Molecular Medicine, Sackler Faculty of Medicine, Tel Aviv University, Ramat Aviv, Tel Aviv 69978, Israel

关键词: Neutral endopeptidase;    Neutral proteinase;    Enkephalinase;    Thermolysin;    Common acute lymphoblastic leukemia antigen;   

DOI  :  10.1016/0014-5793(89)81098-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A sensitive two-stage enzymatic reaction for mammalian and bacterial metalloendopeptidases has been developed using the substrate 3-carboxypropanoyl-alanyl-alanyl-leucine-4-nitroanilide supplemented with Streptomyces griseus amino-peptidase. Neutral endopeptidase EC 3.4.24.11 from bovine kidney hydrolyzes the substrate (pH 7.5, 25°C) with a catalytic efficiency (k _cat=1.2 × 10² s⁻¹, K _m=0. 15 mM) of the highest ever reported for the enzyme acting on synthetic chromophoric and fluorogenic substrates. Thermolysin hydrolyzes the substrate at a faster rate (k _cat=1.2 × 10³ s⁻¹) but the overall efficiency is diminished by a higher K _m (4.2 mM). Suspensions of human neutrophil cells and culture filtrates of Bacillus cereus have been assayed sensitively for their neutral endopeptidase and neutral proteinase activities, respectively. The assay provides a convenient tool for the kinetic investigation of neutral endopeptidases and neutral proteinases and for assessing their function in biological systems.

【 授权许可】

Unknown