title: Ca2+- and Mg2+-dependent association of phosphorylase kinase with human erythrocyte membranes.
abstract: The interaction of rabbit muscle phosphorylase kinase (EC 2.7.1.38) with human erythrocyte membranes was investigated. It was found that at pH 7.0 the kinase binds to the inner face of the erythrocyte membrane (inside-out vesicles) and that this binding is Ca2+- and Mg2+-dependent. The sharpest increase in the binding reaction occurs at concentrations between 70 and 550 nM free Ca2+. Erythrocyte ghost or right-side out erythrocyte vesicles showed a significantly lower capacity to interact with phosphorylase kinase. Autophosphorylated phosphorylase kinase shows a similar Ca2+-dependent binding profile, while trypsin activation of the kinase and calmodulin decrease the original binding capacity by about 50%. Heparin (200 micrograms/ml) and high ionic strength (50 mM NaCl) almost completely blocks enzyme-membrane interaction; glycogen does not affect the interaction.
authors.affiliation: Institute of Biological Research, National Hellenic Research Foundation, Athens, Greece.
authors.forename: S M
authors.initials: SM
authors.lastname: Kyriakidis
authors.forename: T G
authors.initials: TG
authors.lastname: Sotiroudis
authors.forename: A E
authors.initials: AE
authors.lastname: Evangelopoulos
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002147:Calmodulin / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004910:Erythrocyte Membrane / Q000201:enzymology*; D006003:Glycogen / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008274:Magnesium / Q000494:pharmacology*; D009132:Muscles / Q000201:enzymology; D009994:Osmolar Concentration; D010764:Phosphorylase Kinase / Q000097:blood*; D010766:Phosphorylation; D011817:Rabbits; D012965:Sodium Chloride / Q000494:pharmacology; D014357:Trypsin / Q000494:pharmacology
chemical_list: D002147:Calmodulin; D012965:Sodium Chloride; D006493:Heparin; D006003:Glycogen; D010764:Phosphorylase Kinase; D014357:Trypsin; D008274:Magnesium; D002118:Calcium