A SIMPLE METHOD FOR ISOLATING HUMAN ENDOTHELIN-CONVERTING ENZYME FREE FROM CONTAMINATION BY NEUTRAL ENDOPEPTIDASE-24.11

被引：27

作者：

CORDER, R

KHAN, N

HARRISON, VJ

机构：

[1] William Harvey Research Institute, St. Bartholomew’s Hospital Medical College, London EC1M 6BQ, Charterhouse Square

来源：

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS | 1995年 / 207卷 / 01期

基金：

英国惠康基金;

关键词：

D O I：

10.1006/bbrc.1995.1195

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：

Subcellular fractionation of the phosphoramidon sensitive membrane-bound endothelin converting enzyme (ECE-1) activity from homogenates of bovine aortic endothelial cells and the human endothelial cell line EA.hy 926, combined with studies of intact cells, shows ECE-1 to be localised primarily to the plasma membrane with the topology of an ectoenzyme. To overcome the problem of neutral endopeptidase 24.11 contaminating the human ECE-1 activity solubilised from the plasma membrane fractions of EA.hy 926, we have used isoelectric focusing to simultaneously solubilise and separate these activities. The metallopeptidase ECE-1 obtained displayed a neutral pH optimum, a molecular weight of 250 kDa on gel filtration chromatography and was inhibited by phosphoramidon with an IC50 of 0.8 mu M. (C) 1995 Academic Press, Inc.

引用

页码：355 / 362

页数：8

共 16 条

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