Conversion of porcine big endothelin to endothelin by an extract from the porcine aortic endothelial cells.
Conversion of porcine big endothelin (big ET) to endothelin (ET) by an extract from cultured porcine aortic endothelial cells was investigated using a radioimmunoassay (RIA) specific for ET and reverse-phase high performance liquid chromatography (RP-HPLC). When big ET was incubated with the extract at an acid pH in the presence of E-64, a cysteine protease inhibitor, the amount of immunoreactive-ET (IR-ET) in the incubation mixture was greatly increased and the optimum pH for this increased reaction was 4.0. The extract-induced increase in IR-ET was completely inhibited by pepstatin-A. These immunoreactive alterations correlated with those in the vasoconstrictor activity. When the incubation mixture of big ET with the cell extract was applied to the RP-HPLC, the IR-ET eluted at the same retention time as seen with synthetic porcine ET. We suggest that a pepstatin-sensitive aspartic protease is involved in the conversion of big ET to ET in vascular endothelial cells.
Matsumura
Y
Y
Department of Pharmacology, Osaka University of Pharmaceutical Sciences, Japan.
Ikegawa
R
R
Takaoka
M
M
Morimoto
S
S
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D019332:Endothelin-1; D016232:Endothelins; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D009119:Muscle Contraction; D009131:Muscle, Smooth, Vascular / Q000502:physiology; D010436:Pepstatins / Q000378:metabolism; D010455:Peptides / Q000378:metabolism*; D011498:Protein Precursors / Q000378:metabolism*; D011863:Radioimmunoassay; D013552:Swine
D019332:Endothelin-1; D016232:Endothelins; D010436:Pepstatins; D010455:Peptides; D011498:Protein Precursors; C014195:Streptomyces pepsin inhibitor; C031375:pepstatin