title: Proteolysis of protein kinase C: mM and microM calcium-requiring calpains have different abilities to generate, and degrade the free catalytic subunit, protein kinase M.
abstract: Limited proteolysis of protein kinase C (PKC) by calpain under cell free conditions cleaves the regulatory and catalytic PKC subunits, generating a free, co-factor independent catalytic subunit, termed PKM. In the present study, we demonstrate distinct differences in the rate, nature, and lipid-sensitivity of PKC and PKM proteolysis by microM and mM calcium-requiring calpain isozymes (mu calpain or m calpain, respectively). PKC is a preferred substrate for m calpain; not even a 100-fold increase in mu calpain was capable of degrading PKC as fast as in calpain. PKM was generated by both m and mu calpains, but was itself rapidly degraded by m calpain and therefore was only transiently detectable. By contrast, PKM was formed but not degraded by mu calpain, and persisted in the presence of mu calpain long after all PKC had been degraded. Phosphatidyl serine (PS) inhibited PKC hydrolysis by m calpain yet enhanced PKC hydrolysis by mu calpain. The ability of either calpain isoenzyme to degrade [14C]azocasein was unaffected by PS, suggesting that the influence of PS was on PKC conformation. These findings point towards distinct roles for mu and m calpain in PKC regulation.
authors.affiliation: Department of Biological Sciences, University of Massachusetts Lowell 01854, USA.
authors.forename: C M
authors.initials: CM
authors.lastname: Cressman
authors.forename: P S
authors.initials: PS
authors.lastname: Mohan
authors.forename: R A
authors.initials: RA
authors.lastname: Nixon
authors.forename: T B
authors.initials: TB
authors.lastname: Shea
mesh_terms: D000818:Animals; D001921:Brain / Q000201:enzymology; D002118:Calcium / Q000378:metabolism*; D002154:Calpain / Q000378:metabolism*; D002417:Cattle; D004912:Erythrocytes / Q000201:enzymology; D006801:Humans; D011493:Protein Kinase C / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*
chemical_list: D011493:Protein Kinase C; D002154:Calpain; D002118:Calcium