ATP-dependent protease in bovine adrenal cortex. Tissue specificity, subcellular localization, and partial characterization.
Proteolytic activities in bovine adrenocortical mitochondria were investigated using [14C-methyl]casein as a substrate. Washed mitochondria showed a low proteolytic activity at pH 7.5 or 8.2. ATP (5 mM) plus MgCl2 (7.5 mM) stimulated the proteolysis 9 times at pH 8.2. It was further demonstrated unequivocally by various approaches that the ATP-dependent proteolytic activity localizes in mitochondrial matrix. The activity of the solubilized protease was sensitive to N-ethylmaleimide, mersalyl acid, phenylmethylsulfonyl fluoride, o-vanadate, m-vanadate, vanadyl sulfate, and quercetin but not by oligomycin and ouabain. The ATP-dependent proteolytic activity was eluted at the position of 650,000 daltons on an Ultrogel AcA 22 column as a single symmetrical peak. The gel-filtered enzyme showed high specificity to ATP. GTP and UTP partially substituted ATP. ADP, AMP, tripolyphosphate, alpha, beta-methylene ATP, and beta, gamma-methylene ATP had little or no stimulating activity. ATP did not stimulate the activity in the absence of MgCl2. We measured ATP-dependent proteolytic activities in mitochondrial fractions from several tissues in rat and bovine. Adrenal cortex was one of the tissues of highest activity. In addition, we investigated the effect of adrenal atrophy on the ATP-dependent protease activity in rat adrenal. The ATP-dependent protease activity/adrenal decreased by dexamethasone treatment. The extent of the decrease was similar to that of cytochrome oxidase and succinate dehydrogenase, but smaller than that of cytochrome P-450.
Watabe
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S
Kimura
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T
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000302:Adrenal Cortex / Q000187:drug effects / Q000201:enzymology*; D000818:Animals; D002154:Calpain; D002417:Cattle; D002850:Chromatography, Gel; D003907:Dexamethasone / Q000494:pharmacology; D010450:Endopeptidases / Q000378:metabolism; D006160:Guanosine Triphosphate / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D010447:Peptide Hydrolases / Q000378:metabolism*; D051381:Rats; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D014018:Tissue Distribution; D014544:Uridine Triphosphate / Q000378:metabolism
D003907:Dexamethasone; D006160:Guanosine Triphosphate; D000255:Adenosine Triphosphate; D010450:Endopeptidases; D010447:Peptide Hydrolases; D002154:Calpain; D008274:Magnesium; D014544:Uridine Triphosphate