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The fluorescent phospholipid 1-acyl-2-[12-[(7-nitro-2,1,3-benzoxadiazol-4-yl)amino]dodecanoyl]phosphatidylcholine (C12-NBD-PC) was used to study the kinetics of lipid transfer between phospholipid vesicles. A model based on lipid transfer resulting from the diffusion of soluble monomers was found to accurately predict the kinetics of this transfer process.(More)
We measured the nonradiative fluorescence resonance energy transfer between 7-nitro-2,1,3-benzoxadiazol-4-yl (NBD) labeled lipids (amine labeled phosphatidylethanolamine or acyl chain labeled phosphatidylcholine) and rhodamine labeled lipids in large unilamellar dioleoylphosphatidylcholine vesicles. Two new rhodamine labeled lipid analogues, one a(More)
An assay was developed to study the spontaneous transfer and transbilayer movement (flip-flop) of lipid analogs labeled with the fluorescent fatty acid, 5-(5,7-dimethyl BODIPY)-1-pentanoic acid (C5-DMB-) in large unilamellar lipid vesicles comprised of 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC). The assay is based on the concentration-dependent changes(More)
The metabolism and intracellular distribution of a fluorescent analog of phosphatidylinositol (PI), 1,2-[oleoyl,N-(6-[(7-nitrobenz-2-oxa-1,3-diazo-4-yl) aminocaproyl)]-PI (C6-NBD-PI), was examined in monolayer cultures of Swiss 3T3 cells following its insertion into the plasma membrane. Evidence is presented that the exogenously supplied C6-NBD-PI was(More)
Phospholipid transfer between small unilamellar vesicles and Chinese hamster fibroblasts, a process in which cellular uptake of vesicle lipids without concomitant uptake of vesicle contents occurs, was studied. Isotopically asymmetric vesicles were formed by incubation of 3H-labeled dioleoyl phosphatidylcholine or 3H-labeled dioleoyl(More)
A protocol utilizing chemical and enzymatic steps to synthesize several fluorescent (7-nitrobenz-2-oxa-1,3 diazole) analogs of cytidine diphosphate diacylglycerol and phosphatidylinositol in high yields is described. The fluorescent analogs were characterized by phospholipase C digestion, fast atom bombardment mass spectrometry, and HPLC analysis. Studies(More)
We previously identified a novel phospatidylinositol-specific phospholipase C (PI-PLC) present at the surface of Swiss 3T3 cells using a fluorescent analog of PI and showed that this cell surface PI-PLC (csPI-PLC) activity increases with increasing cell density (J. Biol. Chem. 265, 5337-5340 (1990)). Here, we find that csPI-PLC activity also increased in(More)