Localization and biochemical characterization of endothelin-converting enzyme.
We used a variety of biochemical and immunologic techniques to investigate the localization of endothelin-converting enzyme (ECE) in porcine lung, in human umbilical vein endothelial cells (HUVECs), and in the transformed endothelial cell line EA.hy926. Phosphoramidon-sensitive ECE activity was not enriched on immunomagnetically separated angiotensin-converting enzyme (ACE)-bearing luminal endothelial membranes from pig lung, whereas ACE showed a sevenfold enrichment. ECE activity did, however, co-localize with aminopeptidase-N (AP-N) activity on isolated EA.hy926 plasma membranes. Using a monoclonal antibody to rat lung ECE (AEC32-326) and immunofluorescence microscopy, we have demonstrated ECE on the cell surface of endothelial cell lines.
Barnes
K
K
Department of Biochemistry and Molecular Biology, University of Leeds, England.
Murphy
L J
LJ
Takahashi
M
M
Tanzawa
K
K
Turner
A J
AJ
D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000032:analysis / Q000276:immunology / Q000378:metabolism*; D002478:Cells, Cultured; D000072396:Endothelin-Converting Enzymes; D005455:Fluorescent Antibody Technique; D006801:Humans; D008168:Lung / Q000201:enzymology; D008666:Metalloendopeptidases; D051381:Rats; D013552:Swine
D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D000072396:Endothelin-Converting Enzymes