Turner A J, Murphy L J
Department of Biochemistry and Molecular Biology, University of Leeds, U.K.
Biochem Pharmacol. 1996 Jan 26;51(2):91-102. doi: 10.1016/0006-2952(95)02036-5.
A critical processing step in endothelin biosynthesis is the conversion of the intermediate "big endothelin" to its biologically active product catalysed by endothelin converting enzyme (ECE). In this commentary we discuss critically the cellular location, structure, and activity of the isoforms of ECE. The current evidence supporting a metallopeptidase ECE as the physiological regulator of endothelin production is described. Its sensitivity to inhibition by the fungal metabolite phosphoramidon and subsequent cloning of the enzyme indicate it to be a type II integral membrane protein homologous with neural endopeptidase-24.11 (E-24.11), the major neuropeptide-degrading ectoenzyme in brain and other tissues. Unlike E-24.11, however, ECE exists as a disulphide-linked dimer of subunit M(r) 120-130 kDa and is not inhibited by other E-24.11 inhibitors such as thiorphan. Alternative splicing produces two forms of ECE with distinct N-terminal tails. These isoforms of ECE-1 show similar specificity converting big endothelin-1 (ET-1) to ET-1 but big ET-2 and big ET-3 are converted much less efficiently. This suggests that additional forms of ECE remain to be isolated. Immunocytochemical studies indicate a predominant cell-surface location for ECE-1, like E-24.11. This is consistent with the conversion of exogenous big ET-1 when administered in vivo and the inhibition of this event by phosphoramidon. However, mature ET-1 can be detected in intracellular vesicles in endothelial cells, suggesting that some processing occurs in the constitutive secretory pathway. This may be mediated by ECE-2, a recently cloned member of the E-24.11/ECE family which has an acidic pH optimum. Selective inhibitors of ECE may have therapeutic applications in cardiovascular and renal medicine.
内皮素生物合成中的一个关键加工步骤是由内皮素转化酶(ECE)催化中间体“大内皮素”转化为其生物活性产物。在本述评中,我们批判性地讨论了ECE同工型的细胞定位、结构和活性。描述了支持金属肽酶ECE作为内皮素产生的生理调节因子的现有证据。它对真菌代谢产物磷酰胺素抑制作用的敏感性以及随后该酶的克隆表明它是一种与神经内肽酶-24.11(E-24.11)同源的II型整合膜蛋白,E-24.11是脑和其他组织中主要的神经肽降解外切酶。然而,与E-24.11不同,ECE以亚基M(r)120 - 130 kDa的二硫键连接二聚体形式存在,并且不受其他E-24.11抑制剂如硫喷妥的抑制。选择性剪接产生两种具有不同N末端尾巴的ECE形式。这些ECE-1同工型在将大内皮素-1(ET-1)转化为ET-1方面显示出相似的特异性,但大ET-2和大ET-3的转化效率要低得多。这表明仍有待分离其他形式的ECE。免疫细胞化学研究表明,与E-24.11一样,ECE-1主要位于细胞表面。这与体内给予外源性大ET-1时的转化以及磷酰胺素对该事件的抑制作用一致。然而,在内皮细胞的细胞内囊泡中可以检测到成熟的ET-1,这表明在组成型分泌途径中发生了一些加工过程。这可能由ECE-2介导,ECE-2是E-24.11/ECE家族最近克隆的成员,其最适pH为酸性。ECE的选择性抑制剂可能在心血管和肾脏医学中有治疗应用。