Identification and Partial Purification of a Thiol Endothelin- Converting Enzyme from Porcine Aortic Endothelial Cells

Publisher Website
Google Scholar
Abstract
Endothelin is a potent peptide vasoconstrictor. The final step in the processing of endothelin has been postulated to be the cleavage of the Trp21-Va122 peptide bond in proendothelin by a putative endothelin-converting enzyme. A soluble extract of primary porcine aortlc endothelial cells was found to contain an enzyme activity that converted proendothelin-l (proET-1) to an endothelin-1 (ET-1)-like peptide as determined by the rabbit aortlc ring contraction assay. This enzyme was partially purified by DE52 ion-exchange chronuitography. Incubation of proET-1 with the partially purified enzyme generated a product which had a retention time on HPLC Identical to that of authentic ET-1. Further analysis of the product showed that it caused contraction of rabbit aortic rings, had a molecular weight identical to ET-1 as measured by fast atom bombardment mass spectrometry, and competed for [125I]ET-l binding in an RIA using specific antibodies which recognize the carboxy terminal tryptophan of ET-1. The enzyme activity could be inhibited by thiol protease inhibitors such as Z-phe-pheCHN2 and p-hydroxymercuribenzoate, but not by serine- or metalloprotease Inhibitors. The optimal pH for the enzymatic activity was between 7.0 and 7.5, and no activity was detected at pH 4.0. These results demonstrate that this thiol protease is a potential endothelin-converting enzyme.

This publication has 0 references indexed in Scilit: