title: Characterization of the calcium-binding site that regulates association of protein kinase C with phospholipid bilayers.
abstract: Association of calcium-dependent isotypes of protein kinase C (PKC) with a phospholipid bilayer is regulated by a single Ca(2+)-binding site. The dependence of PKC association with phosphatidylserine-containing membranes on the concentration of Ca2+ is linear in the submicro- to submillimolar range. The Ca(2+)-regulated association of PKC with the membrane is sensitive to the factors that alter the diffuse double-layer potential produced by anionic lipids such as phosphatidylserine (PS). This indicates that the Ca(2+)-binding site on the membrane-bound enzyme senses a higher concentration of Ca2+ than is present in bulk solution. This is a consequence of the accumulation of Ca2+ in the layer adjacent to the plane of the membrane by the double-layer potential. Calculations based on the Gouy-Chapman-Stern theory of the diffuse double layer yielded a unique value of the Ca2+ dissociation constant for the Ca(2+)-PKC-bilayer complex equal to approximately 700 nM. The soluble form of the enzyme has a 3.5 order of magnitude lower affinity for Ca2+. The free energy of interaction between the Ca(2+)- and PS-binding sites is large (approximately 5 kcal/mol). In contrast, the interaction between the diacylglycerol-binding site and either the Ca(2+)- or PS-binding site appears to be weak.
authors.affiliation: Department of Biochemistry, McMaster University, Hamilton, Ontario, Canada.
authors.forename: M
authors.initials: M
authors.lastname: Mosior
authors.forename: R M
authors.initials: RM
authors.lastname: Epand
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D008051:Lipid Bilayers / Q000378:metabolism*; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism*; D051381:Rats
chemical_list: D008051:Lipid Bilayers; D010743:Phospholipids; D011493:Protein Kinase C; D002118:Calcium