title: Interaction of protein kinase C with phosphatidylserine. 1. Cooperativity in lipid binding.
abstract: The basis for the apparent cooperativity in the activation of protein kinase C by phosphatidylserine has been addressed using proteolytic sensitivity, resonance energy transfer, and enzymatic activity. We show that binding of protein kinase C to detergent-lipid mixed micelles and model membranes is cooperatively regulated by phosphatidylserine. The sigmoidal dependence on phosphatidylserine for binding is indistinguishable from that observed for the activation of the kinase by this lipid [Newton & Koshland (1989) J. Biol. Chem. 264, 14909-14915]. Thus, protein kinase C activity is linearly related to the amount of phosphatidylserine bound. Furthermore, under conditions where protein kinase C is bound to micelles at all lipid concentrations, activation of the enzyme continues to display a sigmoidal dependence on the phosphatidylserine content of the micelle. This indicates that the apparent cooperativity in binding does not arise because protein kinase C senses a higher concentration of phosphatidylserine once recruited to the micelle. Our results reveal that the affinity of protein kinase C for phosphatidylserine increases as more of this lipid binds, supporting the hypothesis that a domain of phosphatidylserine is cooperatively sequestered around the enzyme.
authors.affiliation: Department of Chemistry, Indiana University, Bloomington 47405.
authors.forename: J W
authors.initials: JW
authors.lastname: Orr
authors.forename: A C
authors.initials: AC
authors.lastname: Newton
mesh_terms: D000818:Animals; D001665:Binding Sites; D004735:Energy Transfer; D004789:Enzyme Activation; D006868:Hydrolysis; D008823:Micelles; D010712:Phosphatidic Acids / Q000737:chemistry; D010718:Phosphatidylserines / Q000737:chemistry*; D010766:Phosphorylation; D011485:Protein Binding; D011493:Protein Kinase C / Q000737:chemistry*; D051381:Rats; D011919:Rats, Inbred Strains; D013329:Structure-Activity Relationship
chemical_list: D008823:Micelles; D010712:Phosphatidic Acids; D010718:Phosphatidylserines; D011493:Protein Kinase C