A microsomal endopeptidase from liver that preferentially degrades stearoyl-CoA desaturase.
A protease was purified some 700-fold from rat liver microsomes by a combination of differential detergent solubilization, hydroxyapatite column chromatography, and gel filtration. The protease exhibits substrate selectivity for stearoyl-CoA desaturase (SCD). The purified protease rapidly degraded SCD while other microsomal proteins including cytochrome b(5) and 11beta-hydroxysteroid dehydrogenase were degraded slowly or not at all. The isolated form of the protease has an apparent molecular mass of approximately 90 kDa. Upon incubation, the 90 kDa form of the protease undergoes rapid conversion to a series of smaller proteins. This conversion is associated with a marked increase in proteolytic activity. Diisopropyl phosphofluoridate (DFP) at high concentration partially inhibited the protease activity. The [(3)H]DFP-labeled protease was detected as three protein bands of approximately 66 kDa under nonreducing conditions and a single 25 kDa band under reducing conditions. The purified protease was inhibited by dithiothreitol, suggesting the presence of an essential disulfide bond. These results further define the mechanism by which SCD is rapidly and selectively degraded in isolated liver microsomes.
Heinemann
F Scott
FS
Department of Pathology, Hoag Memorial Hospital Presbyterian, Newport Beach, California 92663, USA.
Mziaut
Hassan
H
Korza
George
G
Ozols
Juris
J
D000818:Animals; D001055:Apolipoproteins B / Q000378:metabolism; D015786:Cytochromes b5 / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D006913:Hydroxysteroid Dehydrogenases / Q000378:metabolism; D007531:Isoflurophate / Q000494:pharmacology; D008862:Microsomes, Liver / Q000201:enzymology*; D008970:Molecular Weight; D010084:Oxidation-Reduction; D011480:Protease Inhibitors / Q000494:pharmacology; D051381:Rats; D013230:Stearoyl-CoA Desaturase / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D013462:Sulfur Radioisotopes; D014316:Tritium
D001055:Apolipoproteins B; D011480:Protease Inhibitors; D013462:Sulfur Radioisotopes; D014316:Tritium; D007531:Isoflurophate; D015786:Cytochromes b5; D006913:Hydroxysteroid Dehydrogenases; D013230:Stearoyl-CoA Desaturase; D010450:Endopeptidases; D004229:Dithiothreitol