Lipid-dependent membrane enzymes. Kinetic modelling of the activation of protein kinase C by phosphatidylserine.
A previously developed kinetic theory for lipid-dependent membrane enzymes (Sandermann, H. (1982) Eur. J. Biochem. 127, 123-128) is used to examine the activation of protein kinase C by phosphatidylserine. Hill-coefficients ranging up to 11 have been reported for activation in mixed micelles with Triton X-100. On the basis of this uniquely high degree of cooperativity, protein kinase C has been postulated to represent a new class of lipid-dependent membrane enzymes (Newton, A. and Koshland, D.E., Jr. (1989) J. Biol. Chem. 264, 14909-14915). In contrast, activation in the absence of Triton X-100 has led to Hill-coefficients of only less than or equal to 2.6. In order to resolve the apparent discrepancy, activation is now considered to involve binding of PS monomers to interacting sites on the enzyme, a non-activating PS trapping process also occurring in the presence of Triton X-100. Estimates for trapping are made for several sets of published data for micellar activation. The kinetic model developed here successfully fits each data set using a Hill-coefficient of only 3.0. An influence of Ca2+/ions or of a two-step mechanism of lipid-protein interaction are considered as possible molecular explanations. It is concluded (i) that lipid activation of protein kinase C may proceed without unique cooperativity and (ii) that ligand trapping could provide another means for 'threshold-type' kinetic regulation of membrane enzyme and receptor systems.
Institut für Biochemische Pflanzenpathologie, GSF München, Neuherberg, Germany.
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Sandermann
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Duncan
D002118:Calcium / Q000378:metabolism; D003902:Detergents / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D007700:Kinetics; D008024:Ligands; D008563:Membrane Lipids / Q000502:physiology*; D008823:Micelles; D008956:Models, Chemical; D017830:Octoxynol; D010718:Phosphatidylserines / Q000502:physiology*; D011092:Polyethylene Glycols / Q000494:pharmacology; D011493:Protein Kinase C / Q000378:metabolism*
D003902:Detergents; D008024:Ligands; D008563:Membrane Lipids; D008823:Micelles; D010718:Phosphatidylserines; D011092:Polyethylene Glycols; D017830:Octoxynol; D011493:Protein Kinase C; D002118:Calcium