title: A novel method for measuring protein kinase C activity in a native membrane-associated state.
abstract: Physiological activation of protein kinase C (PKC) is believed to occur by redistributing soluble enzyme to the phospholipid environment of membranes. Currently available in vitro methods of measuring PKC activation all involve prior extraction of membrane-associated enzyme and its reconstitution in an artificial phospholipid environment or modification (such as partial trypsinization) of the enzyme itself. Here we report a novel method which, for the first time, allows measurement of active PKC still in its native, membrane-associated state using a specific, physiological substrate. Thus, with this new method PKC activity can be measured while still in an environment that approximates the in vivo situation.
authors.affiliation: Department of Pathology, Faculty of Health Sciences, University of Ottawa, Canada.
authors.forename: B R
authors.initials: BR
authors.lastname: Chakravarthy
authors.forename: D J
authors.initials: DJ
authors.lastname: Franks
authors.forename: J F
authors.initials: JF
authors.lastname: Whitfield
authors.forename: J P
authors.initials: JP
authors.lastname: Durkin
mesh_terms: D000818:Animals; D001839:Bombesin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology; D002462:Cell Membrane / Q000201:enzymology*; D003600:Cytosol / Q000201:enzymology; D004533:Egtazic Acid / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006801:Humans; D008223:Lymphoma / Q000378:metabolism; D051379:Mice; D008970:Molecular Weight; D010750:Phosphoproteins / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats; D013379:Substrate Specificity; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured
chemical_list: D010750:Phosphoproteins; D004533:Egtazic Acid; D004815:Epidermal Growth Factor; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D001839:Bombesin; D002118:Calcium