Purification of a novel endothelin-converting enzyme specific for big endothelin-3

被引：57

作者：

Hasegawa, H

Hiki, K

Sawamura, T

Aoyama, T

Okamoto, Y

Miwa, S ^{[1
]}

Shimohama, S

Kimura, J

Masaki, T

机构：

[1] Kyoto Univ, Fac Med, Dept Pharmacol, Sakyo Ku, Kyoto 6068315, Japan

[2] Kyoto Univ, Fac Med, Dept Neurol, Sakyo Ku, Kyoto 6068315, Japan

[3] Natl Cardiovasc Ctr, Res Inst, Suita, Osaka 5650873, Japan

来源：

FEBS LETTERS | 1998年 / 428卷 / 03期

关键词：

endothelin-3; endothelin-converting enzyme; enzyme purification; bovine iris;

D O I：

10.1016/S0014-5793(98)00554-7

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Endothelin-3 (ET-3), a potent vasoactive peptide, is considered to be produced from big ET-3 by endothelin-converting enzyme (ECE) like the other members of the endothelin family (ET-1 and ET-2). We purified a novel ECE from bovine iris microsomes, The purified enzyme, a 140 kDa protein by SDS-PAGE analysis, converted big ET-3 to ET-3 but not big ET-1, with a K-m value of 0.14 mu M for big ET-3. The conversion to ET-3 nas confirmed with sandwich EEA by monoclonal antibodies, the elution profile of HPLC, and intracellular calcium mobilization in CHO-K1 cells expressing recombinant human ETB receptors, The conversion activity was inhibited by an inhibitor of neutral endopeptidase 23.11 (NEP) phosphoramidon. These results show that ECE-3 purified from bovine iris is a novel metalloprotease totally different from ECE-1 or ECE-2, in that the enzyme is highly specific for big ET-3. (C) 1998 Federation of European Biochemical Societies.

引用

页码：304 / 308

页数：5

共 31 条

[1] TRANSFER OF PROTEINS ACROSS MEMBRANES .2. RECONSTITUTION OF FUNCTIONAL ROUGH MICROSOMES FROM HETEROLOGOUS COMPONENTS [J].