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Palmitoyl oleoyl phosphatidylserine

Manufactured by Avanti Polar Lipids

Palmitoyl-oleoyl-phosphatidylserine is a synthetic phospholipid compound. It serves as a precursor for the production of various phospholipids involved in cellular membrane structure and function.

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3 protocols using palmitoyl oleoyl phosphatidylserine

1

Lipid Bilayer Incorporation of Dengue Virus M Protein

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Example 54

Lipid bilayer studies were performed as described elsewhere (Sunstrom, 1996; Miller, 1986). A lipid mixture of palmitoyl-oleoyl-phosphatidylethanolamine, palmitoyl-oleoyl-phosphatidylserine and palmitoyl-oleoyl-phosphatidylcholine (5:3:2) (Avanti Polar Lipids, Alabaster, Ala.) was used. The lipid mixture was painted onto an aperture of 150-200 μm in the wall of a 1 ml delrin cup. The aperture separates two chambers, cis and trans, both containing salt solutions at different concentrations. The cis chamber was connected to ground and the trans chamber to the input of an Axopatch 200 amplifier. Normally the cis chamber contained 500 mM KCl and the trans 50 mM KCl. The bilayer formation was monitored electrically by the amplitude of the current pulse generated by a current ramp. The potentials were measured in the trans chamber with respect to the cis. The protein was added to the cis chamber and stirred until channel activity was seen. The currents were filtered at 1000 Hz, digitized at 5000 Hz and stored on magnetic disk.

The dengue virus M protein C-terminal peptide (DMVC) was dissolved in 2,2,2-trifluoroethanol (TFE) at 0.05 mg/ml to 1 mg/ml. 10 μl of this was added to the cis chamber of the bilayer which was stirred. Channel activity was seen within 15-30 min.

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2

Lipid Vesicle Preparation by Extrusion

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The appropriate volume of lipid stocks dissolved in chloroform were dried under a nitrogen stream and dried overnight using high vacuum. The dried lipids were re-suspended in 50 mM phosphate buffer, pH 8.0 to a final concentration of 20 mM and large unilamellar vesicles (LUV) were formed by extrusion using a Mini-Extruder (Avanti Polar Lipids, Alabaster, AL). Extrusion was performed using 0.1 μm nucleopore polycarbonate membranes (Whatman, Philadelphia, PA) and the prepared stocks were stored at −4 °C. Lipids used in this study: Palmitoyl-oleoyl-phosphatidylcholine (POPC), palmitoyl-oleoyl-phosphatidylserine (POPS), and 1-palmitoyol-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) were purchased from Avanti Polar Lipids (Alabaster, AL)
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3

Lipid Membrane Fusion Protocol

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Palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidyl-gycerol (POPG), palmitoyloleoylphosphatidylserine (POPS) were obtained from Avanti Polar Lipids (Alabaster, AL). Bovine thrombin was from Fisher Scientific (Pittsburgh, PA). The CLY3 construct containing the fusion of enhanced cyan and enhanced yellow fluorescent proteins was provided by Addgene (Cambridge, MA)[16 (link)]. The mutations were introduced using Site-Directed Mutagenesis Kit (Stratagene, Santa Clara, CA) according to manufacturer’s protocol. 8-aminonaphtalene-1,3,6-trisulfonic acid, disodium salt (ANTS) and p-xylene-bis-pyridinium bromide (DPX) were obtained from Molecular Probes (Eugene, OR).
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