title: The direct measurement of protein kinase C (PKC) activity in isolated membranes using a selective peptide substrate.
abstract: A protein kinase C (PKC)-selective peptide substrate was used to develop a method for measuring PKC activity directly and quantitatively in isolated cell membranes without prior detergent extraction and reconstitution of the enzyme with phosphatidylserine and TPA in the presence of excess Ca2+. This simple and rapid method can reliably measure changes in membrane-associated PKC activity induced by various bioactive compounds such as hormones and growth factors. Also, this method, which measures PKC activity in its native membrane-associated state, has the advantage of being able to distinguish between active and inactive PKC associated with cell membranes.
authors.affiliation: Cell Systems Section, National Research Council of Canada, Ottawa.
authors.forename: B R
authors.initials: BR
authors.lastname: Chakravarthy
authors.forename: A
authors.initials: A
authors.lastname: Bussey
authors.forename: J F
authors.initials: JF
authors.lastname: Whitfield
authors.forename: M
authors.initials: M
authors.lastname: Sikorska
authors.forename: R E
authors.initials: RE
authors.lastname: Williams
authors.forename: J P
authors.initials: JP
authors.lastname: Durkin
mesh_terms: D000595:Amino Acid Sequence; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology*; D007700:Kinetics; D008223:Lymphoma; D008722:Methods; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000032:analysis*; D013379:Substrate Specificity; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D010455:Peptides; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium