Showing papers by "Richard M. Epand published in 1989"

Role of the stereochemistry of the hydroxyl group of cholesterol and the formation of nonbilayer structures in phosphatidylethanolamines

Abstract: The phase behavior of mixtures of cholesterol or epicholesterol with phosphatidylethanolamine was studied by differential scanning calorimetry and by X-ray diffraction. Discrete domains of cholesterol are detected by X-ray diffraction in the L alpha phase of phosphatidylethanolamine from egg yolk and synthetic dielaidoylphosphatidylethanolamine beginning at mole fractions of 0.35-0.4 cholesterol. Separate domains of crystalline epicholesterol can also be detected in the L alpha phase of dielaidoylphosphatidylethanolamine by X-ray diffraction at as little as 0.16 mole fraction of epicholesterol. This is a result of poor miscibility of the epicholesterol with dielaidoylphosphatidylethanolamine. Epicholesterol does not alter the L beta----L alpha transition or bilayer spacing. Epicholesterol also has little effect on the diameter of the cylinders in the hexagonal phase. Formation of the inverted hexagonal phase is facilitated by addition of small amounts of cholesterol (mole fraction less than 0.2) in both egg phosphatidylethanolamine and dielaidoylphosphatidylethanolamine. However, at higher mole fractions of cholesterol, the stability of the liquid-crystalline phase is found to increase markedly for dielaidoylphosphatidylethanolamine but not for egg phosphatidylethanolamine, indicating the importance of the structure of the acyl chains in controlling the relative stability of the lamellar and nonlamellar phases in these systems. In contrast to cholesterol, epicholesterol markedly lowers the L alpha----HII phase transition temperature at low mole fraction of sterol. This result demonstrates the importance of the orientation and motional properties of an additive in determining the L alpha----HII transition temperature.

Dependence of the bilayer to hexagonal phase transition on amphiphile chain length.

Abstract: Several series of amphiphiles of increasing chain length were tested for their abilities to modify the L alpha-HII transition of dielaidoylphosphatidylethanolamine using differential scanning calorimetry. Acylcarnitines, alkyl sulfates, alkylsulfobetaines, and phosphatidylcholines, with chain lengths between about 6 and 12 carbon atoms, show an increasing capacity to raise the L alpha-HII phase transition temperature of phosphatidylethanolamine. This is ascribed to increased partitioning of the added amphiphile from water into the membrane as the chain length increases. Alkyl sulfates and alkyltrimethylammonium bromides have diminished capacities to raise the L alpha-HII transition temperature as the chain length is increased from 12 to 16. This is caused by an increase in the hydrophobic portion of the amphiphile leading to a change in the intrinsic radius of curvature and a decrease in the hydrocarbon packing constraints in the HII phase relative to the shorter chain amphiphiles. The L alpha-HII transition temperature of phosphatidylethanolamine with acylcarnitines of chain length 14-20 carbon atoms, alkylsulfobetaines above 14 carbon atoms, and phosphatidylcholines with acyl groups having above 10 carbon atoms is relatively insensitive to chain length. We suggest that this is caused by a balance between increasing hydrocarbon volume promoting the HII phase through decreased intrinsic radius of curvature and greater relief of hydrocarbon packing constraints vs greater intermolecular interactions favoring the more condensed L alpha phase. This latter effect is more important for amphiphiles with large headgroups which can pack more efficiently in the L alpha phase. The phosphatidylcholines show a gradual decrease in bilayer stabilization between 10 and 22 carbon atoms.(ABSTRACT TRUNCATED AT 250 WORDS)

Studies on the mechanism of action of a bilayer stabilizing inhibitor of protein kinase C: cholesterylphosphoryldimethylethanolamine.

Abstract: Cholesterylphosphoryldimethylethanolamine is a zwitterionic compound which is a good bilayer stabilizer As has been found with many other compounds having these properties, cholesterylphosphoryldimethylethanolamine is found to be a potent inhibitor of protein kinase C in both vesicle and micelle assay systems The kinetics of the inhibition in Triton X-100 micelles was non-competitive with respect to ATP, histone, diolein, phorbol ester and Ca2+ It has a Ki of about 30 μm The inhibition kinetics as a function of phosphatidylserine concentration is more complex but suggestive of competitive inhibition Cholesterylphosphoryldimethylethanolamine does not prevent the partitioning of protein kinase C into the membrane This inhibitor lowers the Ca2+-phosphatidylserine-independent phosphorylation of protamine sulfate by protein kinase C and directly affects the catalytic segment of the enzyme generated by tryptic hydrolysis Thus, this zwitterionic bilayer stabilizing inhibitor of protein kinase C both competes with the binding of phosphatidylserine as well as affects the active site of protein kinase C

Potent inhibitors of glucagon-stimulated adenylate cyclase associated with serum lipoprotein particles.

Abstract: 1, v/v). The inhibition was greater against glucagon-stimulated activity than against basal activity. Acetone extraction increased the potency of inhibition. All three lipoprotein fractions, i.e., very low, low, and high density lipoproteins, released some inhibitory component after acetone extraction. The inhibitor was concentrated in the lipoprotein fractions, since acetone extraction of plasma did not release an inhibitor. The acetone extract from the very low density liproprotein was the most inhibitory. This material was further purified and partially characterized. The inhibitor had a molecular mass of about 500. It was inhibitory at micromolar concentrations. The material was sufficiently hydrophobic to migrate in normal-phase thin-layer chromatography (TLC). Nuclear magnetic resonance results indicated that it was not a polar lipid. There were several different inhibitory factors that were separable by TLC. The sequestration of these inhibitors into lipoproteins reduced their effectiveness in inhibiting the action of counter-regulatory hormones, such as glucagon. Key words: glucagon, adenylate cyclase, lipoprotein, diabetes mellitus. BILAN, P. J., WEITZMAN, J. B., PFLIEGER, T., LANCASTER, C. R. D., STAFFORD, A., et EPAND R. M. 1989. Potent inhibitors of glucagon-stimulated adenylate cyclase associated with serum lipoprotein particles. Biochem. Cell Biol.