A permanent human cell line (EA.hy926) preserves the characteristics of endothelin converting enzyme from primary human umbilical vein endothelial cells.
Purification of endothelin converting enzyme (ECE) from endothelial cells has been hindered by the difficulty in obtaining primary endothelial cells in large quantity. We therefore tested transformed human umbilical vein endothelial cells (EA.hy926) for ECE activity. Our data clearly demonstrate that this transformed cell line preserves the ECE properties of the primary cell line. These include: (i) one sharp activity optimum at neutral pH; (ii) characteristics typical of a metalloprotease; (iii) IC50 value for phosphoramidon of 1.8 microM (2.7 microM for HUVEC); (iv) no inhibition by captopril and thiorphan, inhibitors of angiotensin converting enzyme and neutral endopeptidase 24.11. The enzyme showed a substrate specificity for big ET-1:big ET-2:big ET-3 in a ratio of 40:2.5:1. This report presents evidence that a permanent human endothelial cell line, EA.hy926, preserves the ECE activity of HUVEC and is useful for the study of ECE and its regulation of ET-1 production.
Ahn
K
K
Department of Biochemistry, Parke-Davis Pharmaceutical Research, Division of Warner-Lambert Company, Ann Arbor, Michigan 48105, USA.
Pan
S
S
Beningo
K
K
Hupe
D
D
D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D002417:Cattle; D002461:Cell Line, Transformed / Q000201:enzymology*; D000072396:Endothelin-Converting Enzymes; D016232:Endothelins / Q000378:metabolism; D004730:Endothelium, Vascular / Q000201:enzymology*; D006020:Glycopeptides / Q000378:metabolism; D006801:Humans; D008666:Metalloendopeptidases; D015260:Neprilysin / Q000378:metabolism; D013379:Substrate Specificity; D014471:Umbilical Veins / Q000166:cytology; D015032:Zinc / Q000494:pharmacology
D016232:Endothelins; D006020:Glycopeptides; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D015260:Neprilysin; D000072396:Endothelin-Converting Enzymes; D015032:Zinc; C008890:phosphoramidon