Purification and Characterization of Three Types of Protein Kinase C from Rabbit Brain Cytosol

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 84; no. 13; pp. 4418 - 4422
• Main Authors: Jaken, Susan, Kiley, Susan C.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 01.07.1987; National Acad Sciences
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Antibodies; Antibodies, Monoclonal - immunology; Biochemistry; Biological and medical sciences; Brain Chemistry; Caenorhabditis elegans Proteins; Calcium - metabolism; Carrier Proteins; Chromatography; Chromatography, Affinity; Cytosol; Cytosol - enzymology; Enzymes and enzyme inhibitors; Fundamental and applied biological sciences. Psychology; Histones; Isoenzymes - immunology; Isoenzymes - isolation & purification; Isoenzymes - metabolism; Legends; Molecular Weight; Phorbol 12,13-Dibutyrate; Phorbol esters; Phorbol Esters - metabolism; Phosphatidylserines - metabolism; Protein isoforms; Protein Kinase C - immunology; Protein Kinase C - isolation & purification; Protein Kinase C - metabolism; Rabbits; Receptors, Drug; Receptors, Immunologic - isolation & purification; Receptors, Immunologic - metabolism; Substrate Specificity; Transferases; Ungulates; Characterization; Vertebrata; Molecular form; Protein kinase C; Mammalia; Purification; Isozyme; Central nervous system; Rabbit; Lagomorpha; Cytosol
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.84.13.4418

Three types of protein kinase C were purified from rabbit brain cytosol. Each type has a molecular mass of ≈ 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.