Role of mast cell chymase in the extracellular processing of big-endothelin-1 to endothelin-1 in the perfused rat lung.
Previous studies of endothelin-1 (ET) synthesis have shown that some cultured endothelial cells secrete an intermediate product, big-endothelin-1 (bigET), suggesting that the processing of secreted bigET to ET may be physiologically significant. In this study, two pertinent ET converting enzyme activities, mast cell chymase I (EC 3.4.21.39) and a phosphoramidon-sensitive, neutral metalloprotease, were identified in a rat lung particulate fraction. We perfused rat lungs with bigET and chymostatin or phosphoramidon to study the relevance of these two proteases to the processing of extracellular bigET in vivo. Addition of compound 48/80 (a compound which activates mast cells, causing degranulation and release of chymase) to the perfusion buffer greatly increased hydrolysis of exogenously added bigET to ET. ET formation was inhibited completely by 32 microM chymostatin, whereas inhibition by 50 microM phosphoramidon was incomplete and variable. Perfusate histamine levels were used to monitor the extent of mast cell degranulation, and inhibition of ET production by phosphoramidon was attributed to inhibition of degranulation, per se. There was a direct correlation between perfusate ET and histamine levels in both control and phosphoramidon-treated (but not chymostatin-treated) lungs. Our results suggest that chymase from lung mast cells is capable of physiologically relevant extracellular processing by bigET to ET in the perfused rat lung.
Wypij
D M
DM
Glaxo Research Institute, Research Triangle Park, NC 27709.
Nichols
J S
JS
Novak
P J
PJ
Stacy
D L
DL
Berman
J
J
Wiseman
J S
JS
D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000037:antagonists & inhibitors; D053818:Chymases; D019332:Endothelin-1; D000072396:Endothelin-Converting Enzymes; D016232:Endothelins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006020:Glycopeptides / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D008168:Lung / Q000201:enzymology*; D008407:Mast Cells / Q000201:enzymology*; D008666:Metalloendopeptidases; D009842:Oligopeptides; D010477:Perfusion; D011498:Protein Precursors / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012697:Serine Endopeptidases / Q000378:metabolism*
D019332:Endothelin-1; D016232:Endothelins; D006020:Glycopeptides; D009842:Oligopeptides; D011498:Protein Precursors; D012697:Serine Endopeptidases; D053818:Chymases; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D000072396:Endothelin-Converting Enzymes; C008890:phosphoramidon