Purification and characterization of three types of protein kinase C from rabbit brain cytosol.
Three types of protein kinase C were purified from rabbit brain cytosol. Each type has a molecular mass of approximately 80 kDa and serves as a receptor for phorbol esters. Polyclonal antibodies produced to two protein kinase C types were relatively type-specific, indicating that these proteins have unique antigenic determinants. We, therefore, characterized the enzymatic activities to determine if these proteins also had distinct biochemical properties. Type 1 protein kinase C was relatively less Ca2+-dependent than types 2 and 3. The addition of Ca2+ increased Vmax approximately 40% for type 1,600% for type 2, and 1400% for type 3 as compared to the Vmax measured at lower Ca2+ conditions. These results suggest that differences in primary structure can confer type-specific biochemical properties, and this in turn may provide the basis for protein kinase C type-specific stimulus-response coupling.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001923:Brain Chemistry; D029742:Caenorhabditis elegans Proteins; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins; D002846:Chromatography, Affinity; D003600:Cytosol / Q000201:enzymology; D007527:Isoenzymes / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D008970:Molecular Weight; D015240:Phorbol 12,13-Dibutyrate; D010703:Phorbol Esters / Q000378:metabolism; D010718:Phosphatidylserines / Q000378:metabolism; D011493:Protein Kinase C / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D011817:Rabbits; D011955:Receptors, Drug; D011971:Receptors, Immunologic / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity
D000911:Antibodies, Monoclonal; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D007527:Isoenzymes; D010703:Phorbol Esters; D010718:Phosphatidylserines; D011955:Receptors, Drug; D011971:Receptors, Immunologic; C083141:phorbol ester binding protein; C027776:phorbol ester receptor; D015240:Phorbol 12,13-Dibutyrate; D011493:Protein Kinase C; D002118:Calcium