Document Detail

N-ethylmaleimide differentiates endothelin converting activity by two types of metalloproteinases derived from vascular endothelial cells.
MedLine Citation:
PMID:  1859414     Owner:  NLM     Status:  MEDLINE    
We have recently found that cultured vascular endothelial cells (ECs) contain two types of metalloproteinases which convert big endothelin-1 (big ET-1) to endothelin-1 (ET-1) via a single cleavage between Trp21 and Val22. In the present study, two enzymes were clearly differentiated by using sulfhydryl blocking reagents and anion-exchange HPLC. As reported, the converting activity of the membrane fraction of ECs was specifically inhibited by phosphoramidon. N-ethylmaleimide (NEM) markedly enhanced the apparent converting activity of the membrane fraction. This enhancement was not due to the direct action on the converting enzyme, but rather to inhibition of the degradation of big ET-1 and/or ET-1. In contrast, the converting activity of the cytosolic fraction was abolished by NEM treatment. Effects of phosphoramidon and NEM on converting activities of both fractions were confirmed after anion-exchange HPLC of each fraction, using a COSMOGEL QA column. Our results provide new information on two types of metalloproteinases which convert big ET-1 to ET-1, in vascular ECs.
Y Matsumura; R Ikegawa; Y Tsukahara; M Takaoka; S Morimoto
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biochemical and biophysical research communications     Volume:  178     ISSN:  0006-291X     ISO Abbreviation:  Biochem. Biophys. Res. Commun.     Publication Date:  1991 Jul 
Date Detail:
Created Date:  1991-08-28     Completed Date:  1991-08-28     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0372516     Medline TA:  Biochem Biophys Res Commun     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  531-8     Citation Subset:  IM    
Department of Pharmacology, Osaka University of Pharmaceutical Sciences, Japan.
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MeSH Terms
Aorta, Thoracic
Cell Membrane / enzymology
Cells, Cultured
Chromatography, High Pressure Liquid
Cytosol / enzymology
Endothelins / biosynthesis*,  genetics,  metabolism*
Ethylmaleimide / pharmacology*
Isoenzymes / isolation & purification,  metabolism*
Metalloendopeptidases / isolation & purification,  metabolism*
Muscle, Smooth, Vascular / enzymology*
Protein Precursors / genetics,  metabolism*
Protein Processing, Post-Translational
Reg. No./Substance:
0/Endothelin-1; 0/Endothelins; 0/Isoenzymes; 0/Protein Precursors; 128-53-0/Ethylmaleimide; EC 3.4.24.-/Metalloendopeptidases

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