A calmodulin endoproteinase from mitochondrial membranes.
A proteinase specific for calmodulin has been identified in a crude rat kidney Triton-extracted or sonicated mitochondrial fraction and solubilized by EGTA extraction of these membranes. Mitochondrial fractions from other tissues had less activity, with relative activities: kidney = spleen greater than testes greater than liver, and no detectable activity in either brain or skeletal muscle. This enzyme is active in the presence of EGTA, but not in the presence of calcium, and cleaves calmodulin into three major peptide fragments with Mr 6000, 9000 and 10,000. N-methylated and non-methylated calmodulins were both cleaved by calmodulin proteinase and while troponin was a poor substrate, it was cleaved in the presence of either calcium or EGTA. No other EF hand calcium-binding proteins or other major mitochondrial proteins were cleaved by this enzyme. The peptides resulting from calmodulin proteinase action were isolated by reverse-phase high performance liquid chromatography (HPLC) and sequenced. Sequence analysis indicated that calmodulin proteinase cleaves calmodulin at Lys-75. The effects of proteinase inhibitors indicate that calmodulin proteinase is a trypsin-like enzyme belonging to the serine endopeptidase family of enzymes.
Wright
L S
LS
Department of Pediatrics, University of Wisconsin Medical School, Madison.
Collins
J H
JH
Finn
K A
KA
Siegel
F L
FL
D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium; D002147:Calmodulin / Q000302:isolation & purification*; D002412:Cations; D002851:Chromatography, High Pressure Liquid; D004533:Egtazic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000302:isolation & purification* / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000201:enzymology*; D007668:Kidney / Q000201:enzymology*; D008928:Mitochondria / Q000201:enzymology*; D010089:Oxidoreductases, N-Demethylating / Q000378:metabolism; D010455:Peptides / Q000302:isolation & purification; D051381:Rats; D012697:Serine Endopeptidases; D013379:Substrate Specificity; D014336:Troponin / Q000378:metabolism
D002147:Calmodulin; D002412:Cations; D006657:Histones; D010455:Peptides; D014336:Troponin; D004533:Egtazic Acid; D010089:Oxidoreductases, N-Demethylating; D010450:Endopeptidases; D012697:Serine Endopeptidases; D002118:Calcium