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
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DOI：
10.1016/s0014-5793(03)00310-7
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发表时间：
2003-04-24
期刊：
影响因子：
3.5
通讯作者：
Ferro, ES
中科院分区：
文献类型：
--
作者：
Oliveira, V;Araújo, MC;Ferro, ES
Neurolysin（EP 24.16）和thimet oligopeptidase（EP 24.15）是密切相关的金属内肽酶。将Tyr（613）（EP 24.16）或Tyr（612）（EP 24.15）定点突变为Phe或Ala促进了两种酶的k（cat）/K-M的强烈降低。这些数据表明，在底物水解过程中，这些肽酶的羟基和芳环在这个特定的位置的重要性。此外，EP 24.15 A607 G突变体显示出对于Abz-GFSIFRQ-EDDnp底物的k（cat）/K-M为2 × 10（5）M-1 s（-1），类似于在相应位置含有Gly的EP 24.16（k（cat）/K-M = 3 × 10（5）M-1 s（-1））;野生型EP 24.15对该底物的k（cat）/K-M为2.5 × 10（4）M-1 s（-1）。(C)2003年欧洲生物化学学会联合会。由Elsevier Science B. V.出版，版权所有。
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.