diphenylhexatriene and stearic-acid

Membrane proteins, such as viral spike, were transferred spontaneously from influenza virus-infected cells to various liposomes. The protein transfer was enhanced by the presence of negative charged component dicetylphosphate (DCP) or stearic acid (SA) in dimyristoylphosphatidylcholine (DMPC) liposomes. The lowering of membrane fluidity did not relate to the effect of DCP or SA on protein transfer in this study. We considered that the alteration of membrane properties, such as construction of the surface or stability of transferred protein in liposomes, due to the specific structure of DCP or SA is responsible for the enhancement of spontaneous protein transfer by the presence of the amphiphilic components.

Topics: Animals; Cell Line; Dimyristoylphosphatidylcholine; Diphenylhexatriene; Electrophysiology; Fluorescence Polarization; Fluorescent Dyes; Fluorometry; Haplorhini; Kidney; Liposomes; Membrane Fluidity; Organophosphates; Orthomyxoviridae; Stearic Acids; Viral Matrix Proteins

To investigate NADPH-oxidase activation, we studied the effects of free fatty acid (FFA), their uncharged derivatives, and calcium on membrane lipid structure and superoxide anion (O2-) release from intact neutrophils and in cell-free O2(-)-generating systems. This study determined that in calcium-free phosphate-buffered saline (PBS), cis-unsaturated FFA (cis FFA), trans-unsaturated FFA (trans FFA), and to a limited extent saturated FFA decreased the polarization of the membrane lipid structure probe 1,6-diphenyl-1,3,5-hexatriene (DPH), and these decreases correlated with partitioning of the FFA into the plasmalemma and the release of O2- from intact neutrophils and in cell-free preparations. Although a decrease in DPH polarization was always observed under conditions that resulted in the release of O2-, there was not a direct correlation between the amount of decrease in DPH polarization and the release of O2-. Trans FFA did not induce a dose-dependent decrease or as dramatic a decrease in DPH polarization compared with cis FFA, yet the trans FFA stimulated a greater release of O2- at 2.5, 5, and 10 microM concentrations. In addition, responses of the neutrophil to trans FFA, but not cis FFA, were differentially affected by the presence of calcium. When 0.1 mM calcium was added to the PBS decreases in DPH polarization in response to trans FFA were reduced by greater than 60%, whereas O2- was reduced by only 25-36%. The addition of 0.1 mM calcium 3 min after the trans FFA had partitioned into the membrane also reversed by 50-65% decreases in DPH polarization but did not affect the release of O2-. In the presence of 0.9 mM calcium, only the cis FFA decreased DPH polarization or stimulated the release of O2-. Calcium is known to interact more readily with the ionized carboxyl group of trans FFA, neutralizing the anionic charge through an electrostatic interaction. In support of the requirement for the ionized carboxyl group, structurally similar uncharged cis, trans, and saturated fatty alcohols; methyl esters; and aldehydes decreased DPH polarization in the absence and presence of 0.9 mM calcium; however, none of these compounds stimulated the release of O2-. These results indicate that, in addition to the partitioning of FFA into the plasmalemma, which results in calcium-modulatable decreases in DPH polarization, activation or assembly of the NADPH-oxidase requires the ionized carboxyl group.

Topics: Animals; Arachidonic Acid; Arachidonic Acids; Calcium; Cell Membrane; Cricetinae; Diphenylhexatriene; Dose-Response Relationship, Drug; Eicosanoic Acids; Fatty Acids, Nonesterified; Linolenic Acids; Membrane Lipids; Myristic Acid; Myristic Acids; NADH, NADPH Oxidoreductases; NADPH Oxidases; Neutrophils; Oleic Acid; Oleic Acids; Stearic Acids; Superoxides

Erythrocyte membranes and their liposomes were prepared from clinically normal dogs and Labrador retrievers with hereditary muscular dystrophy. The "static" and "dynamic" components of fluidity of each membrane were then assessed by steady-state fluorescence polarization techniques using limiting hindered fluorescence anisotropy and order parameter values of 1,6-diphenyl-1,3,5-hexatriene (DPH) and fluorescence anisotropy values of DL-2-(9-anthroyl)-stearic acid and DL-12-(9-anthroyl)-stearic acid, respectively. Membrane lipids were extracted and analyzed by thin-layer chromatography and gas chromatography. The results of these studies demonstrated that the lipid fluidity of erythrocyte membranes, and their liposomes, prepared from dystrophic dogs were found to possess significantly lower "static and dynamic components of fluidity" than control counterparts. Analysis of the composition of membranes from dystrophic dogs revealed a higher ratio of saturated fatty acyl chain/unsaturated chains (w/w) and lower double-bond index. Alterations in the fatty acid composition such as decrease in levels of linoleic (18:2) and arachidonic (20:4) acids and increase in palmitic (16:0) and stearic (18:0) acids were also observed in the membranes of dystrophic animals. These associated fatty acyl alterations could explain, at least in part, the differences in membrane fluidity between dystrophic and control dogs.

Topics: Animals; Arachidonic Acid; Arachidonic Acids; Diphenylhexatriene; Dogs; Erythrocyte Membrane; Fluorescence Polarization; Fluorescent Dyes; Linoleic Acid; Linoleic Acids; Membrane Fluidity; Membrane Lipids; Muscular Dystrophy, Animal; Palmitic Acid; Palmitic Acids; Stearic Acids

Thymocytes, obtained from young rats, were incubated in the presence of either diheptadecanoylphosphatidylcholine or dioleylphosphatidylcholine vesicles and desaturation of either [1-14C]stearic acid or [1-14C]linoleic acid was followed in the endoplasmic reticulum. Incubation with diheptadecanoylphosphatidylcholine resulted in an accumulation of heptadecanoic acid in the plasma membrane, but not in the endoplasmic reticulum and mitochondria, and an increase in membrane ordering as assessed by diphenylhexatriene fluorescence polarization. A shift to higher temperature of the phase separation in the plasma membrane was also observed. Both delta 9 and delta 6 desaturase activities were enhanced in these cells, with delta 6 responding more intensly. Accumulation of oleic acid in the plasma membrane could not be observed when the cells were incubated in the presence of dioleylphosphatidylcholine vesicles, but all the membranes separated, including the microsomes, became more fluid. This can be attributed to removal of cholesterol by the vesicles. Fluidization of plasma membrane and endoplasmic reticulum depressed the conversion of stearate to oleate and linoleate to gamma-linolenate. It is concluded that there is an exchange of information between the plasma membrane and the endoplasmic reticulum in order to maintain the proper fluidity relationships and that this occurs without transfer of lipids from the former to the latter.

Topics: Animals; Cell Membrane; Diphenylhexatriene; Endoplasmic Reticulum; Fatty Acid Desaturases; Fatty Acids; Fluorescent Dyes; Intracellular Membranes; Linoleic Acid; Linoleic Acids; Male; Membrane Fluidity; Microsomes; Mitochondria; Rats; Stearic Acids; T-Lymphocytes