Shimada K, Takahashi M, Tanzawa K
Biological Research Laboratories, Sankyo Co., Ltd., Tokyo, Japan.
J Biol Chem. 1994 Jul 15;269(28):18275-8.
Endothelin (ET) is a 21-residue potent vasoconstrictive peptide produced by vascular endothelial cells and formed from its precursor, big endothelin (big ET), by endothelin-converting enzyme (ECE). Here we report the cloning and functional expression of a complementary DNA encoding a rat ECE from endothelial cells. Rat ECE is a highly glycosylated protein consisting of 10 possible N-linked glycosylation sites, a zinc-binding domain, and a single membrane-spanning region. It has structural and sequence homology to neutral endopeptidase and Kell blood group protein, a putative neutral endopeptidase. Monoclonal antibody risen against purified rat lung ECE recognized broad glycosylated bands in membrane fractions prepared from both rat lung and COS cells transfected with a rat ECE expression vector. Expressed ECE was inhibited by phosphoramidon, but not by thiorphan. It also cleaved big ET-1 efficiently but not big ET-2 or big ET-3. Northern blot analysis revealed that ECE messenger RNA is widely expressed in many tissues.
内皮素（ET）是一种由血管内皮细胞产生的含21个氨基酸残基的强效血管收缩肽，它由其前体大内皮素（big ET）通过内皮素转换酶（ECE）形成。在此，我们报告了从内皮细胞中克隆并功能性表达编码大鼠ECE的互补DNA。大鼠ECE是一种高度糖基化的蛋白质，包含10个可能的N-连接糖基化位点、一个锌结合结构域和一个单一跨膜区域。它与中性内肽酶和凯尔血型蛋白（一种假定的中性内肽酶）具有结构和序列同源性。针对纯化的大鼠肺ECE产生的单克隆抗体在从大鼠肺和用大鼠ECE表达载体转染的COS细胞制备的膜组分中识别出广泛的糖基化条带。表达的ECE受到磷酰胺脒的抑制，但不受硫磷酰胺的抑制。它还能有效切割大ET-1，但不能切割大ET-2或大ET-3。Northern印迹分析显示ECE信使RNA在许多组织中广泛表达。
