Showing papers by "Richard M. Epand published in 1991"

Evidence for the regulation of the activity of protein kinase C through changes in membrane properties.

Abstract: We measured the effects of two branched-chain analogs of distearoyl-phosphatidylcholine, containing either a methyl or an n-butyl group at the 8 position, on the bilayer to hexagonal phase transition temperature of dielaidoylphosphatidylethanolamine. The former compound raised the bilayer to hexagonal phase transition temperature while the latter compound lowered it. The opposite effects of these amphiphiles on protein kinase C activity (inhibition and activation, respectively) correlated with their effects on lipid polymorphism. Because of the similarity of the structures of these two compounds, it seems likely that their opposite effects on the activity of protein kinase C is a result of their alteration of the lipid environment of the membrane rather than to binding to a specific site on the protein.

Mimicking the membrane-mediated conformation of dynorphin A-(1-13)-peptide: circular dichroism and nuclear magnetic resonance studies in methanolic solution.

Abstract: The structural requirements for the binding of dynorphin to the {kappa}-opioid receptor are of profound clinical interest in the search for a powerful nonaddictive analgesic. These requirements are thought to be met by the membrane-mediated conformation of the opioid peptide dynorphin A-(1-13)-peptide, Tyr{sup 1}-Gly{sup 2}-Gly{sup 3}-Phe{sup 4}-Leu{sup 5}-Arg{sup 6}-Arg{sup 7}-Ile{sup 8}-Arg{sup 9}-Pro{sup 10}-Lys{sup 11}-Leu{sup 12}-Lys{sup 13}. Schwyzer has proposed an essentially {alpha}-helical membrane-mediated conformation of the 13 amino acid peptide. In the present study, circular dichroism (CD) studies on dynorphin A-(1-13)-peptide bound to an anionic phospholipid signified negligible helical content of the peptide. CD studies also demonstrated that the aqueous-membraneous interphase may be mimicked by methanol. The 500- and 620-MHz {sup 1}H nuclear magnetic resonance (NMR) spectra of dynorphin A-(1-13)-peptide in methanolic solution were sequence-specifically assigned with the aid of correlated spectroscopy (COSY), double-quantum filtered phase-sensitive COSY (DQF-COSY), relayed COSY (RELAY), and nuclear Overhauser enhancement spectroscopy (NOESY). 2-D CAMELSPIN/ROESY experiments indicated that at least the part of the molecule from Arg{sup 7} to Arg{sup 9} was in an extended or {beta}-strand conformation, which agreed with deuterium-exchange and temperature-dependence studies of the amide protons and analysis of the vicinal spin-spin coupling constants {sup 3}J{sub HN{alpha}}. The results clearly demonstrated the absencemore » of extensive {alpha}-helix formation. {chi}{sub 1} rotamer analysis of the {sup 3}J{sub {alpha}{beta}} demonstrated no preferred side-chain conformations.« less

Reversal of intrinsic multidrug resistance in Chinese hamster ovary cells by amiloride analogs.

Abstract: A number of amiloride analogs can sensitise wild type Chinese Hamster ovary (CHO) cells to the cytotoxic action of vinblastine, daunomycin, puromycin or colchicine. Some of these analogs also have weak sensitising effects on the multidrug resistant CHO cell line, CHRC5. The unusual feature of most of the active amiloride analogs is that they are more potent in reversing the intrinsic multidrug resistance (MDR) phenotype of CHO cells than their acquired MDR characteristic. Human HeLa cells that do not exhibit intrinsic MDR are not affected by these agents. Several of the amiloride analogs have a greater effect in increasing adriamycin uptake in wild type CHO cells than they do with CHRC5 cells. The differential effect of amiloride analogs on intrinsic versus acquired MDR characteristics of Chinese hamster cells suggests some differences in the underlying resistance mechanisms.

Action of insulin in rat adipocytes and membrane properties.

Abstract: Several small peptides inhibit insulin-promoted glucose uptake in rat adipocytes. At 10 {mu}M peptide concentration, the extent of their inhibition of the insulin effect is related to the ability of these peptides to raise the bilayer- to hexagonal-phase transition temperature in model membranes. Hexane and DL-threo-dihydrosphingosine lower this phase transition temperature in model membranes, and they promote glucose uptake in adipocytes. There is thus an empirical relationship between the action of membrane additives on glucose uptake in adipocytes and their effect on the hexagonal-phase-forming tendency in model membranes. The most potent of the bilayer-stabilizing peptides tested in this work is carbobenzoxy-D-Phe-L-Phe-Gly. This peptide also inhibits insulin-stimulated protein synthesis in adipocytes. In contrast, DL-threo-dihydrosphingosine stimulates protein synthesis. The uptake of ({sup 125}I)iodinsulin by adipocytes is inhibited by carbobenzoxy-D-Phe-L-Phe-Gly. The mechanism of action of the bilayer-stabilizing peptides includes inhibition of insulin-dependent protein phosphorylation in adipocytes. The peptides are not specific inhibitors of a single function but are suggested to cause their effects by altering the physical properties of the membrane in a nonspecific manner. These results demonstrate that insulin-dependent functions of rat adipocytes can be modified by membrane additives in a manner predictable from the properties of these additives in model membranes.

Biological Consequences of Alterations in the Physical Properties of Membranes

Abstract: Membranes serve important functions for cell organization and for the transduction of signals from the external environment to the cell interior. These functions are generally determined by specific molecules in the membrane such as receptors and membrane bound enzymes. The functioning of these receptors and enzymes are modulated by the nature of their physical environment, i.e. the efficiency of signal transduction and the activity of membrane-bound enzymes will be affected by the nature of the membrane surrounding the specific functional sites. In addition, some properties of membranes such as permeability or membrane fusion may not be absolutely dependent on the presence of specific proteins but may also occur by non-specific mechanisms. In this review, we will focus on the modulation of viral fusion, protein kinase C activity and insulin signalling in adipocytes as examples of membrane functions that are modulated by the physical properties of the membrane. An earlier review of our work in this area has recently appeared (Epand, 1990a).