1 palmitoyl 2 oleoyl sn glycero 3 phosphatidylserine

Manufactured by Avanti Polar Lipids

Sourced in United States

1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylserine is a phospholipid compound. It serves as a structural component of biological membranes.

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Palmitoyl-oleoyl-phosphatidylserine Phosphatidylserine

8 protocols using 1 palmitoyl 2 oleoyl sn glycero 3 phosphatidylserine

Reconstitution and Characterization of αABc Complex

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Cholesterol (Chol), sphingomyelin (SM), and phospholipids (PLs): 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylCholine (POPC), 1-palmditoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylserine (POPS), were obtained dissolved in chloroform from Avanti Polar Lipids, Inc. (Alabaster, AL, USA). Cholesterol analog cholestane spin label (CSL), HEPES, Tris-HCl, NaN₃, sodium chloride (NaCl) lysozyme, deoxycholic acid, and DNase I were obtained from Sigma Aldrich (St. Louis, MO, USA). Isopropyl-1-thio-β-D-galactopyranoside (IPTG), ampicillin, phenylmethylsulfonyl fluoride, and polyethyleneimine were obtained from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA). Recombinant human αAc and αBc were expressed and purified, and the reconstituted 3:1 heteromeric complex of αAc to αBc (i.e., αABc) was prepared using the methods described in Section 4.2 and stored in HEPES buffer (10 mM HEPES, 100 mM NaCl, pH = 7.4). All preparations of α-crystallin (αAc, αBc, and αABc) and the Chol/MHLL membranes, as well as associated binding studies, were performed in HEPES buffer (10 mM HEPES, 100 mM NaCl, pH = 7.4).

Timsina R., Hazen P., Trossi-Torres G., Khadka N.K., Kalkat N, & Mainali L. (2024). Cholesterol Content Regulates the Interaction of αA-, αB-, and α-Crystallin with the Model of Human Lens-Lipid Membranes. International Journal of Molecular Sciences, 25(3), 1923.

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Lipid Membrane Composition and Probes

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1-Palmitoyl-2-oleoyl-sn-glycero-3-PC (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-PE (POPE), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylserine (POPS), cardiolipin, and brain PI(4,5)P₂ were purchased from Avanti Polar Lipids. Rhodamine DHPE (L-1392) and DPH were from Thermo Fisher Scientific. MitoTracker green FM and Red CMXRos were from Thermo Fisher Scientific.

Wang L., Yan Z., Vihinen H., Eriksson O., Wang W., Soliymani R., Lu Y., Xue Y., Jokitalo E., Li J, & Zhao H. (2019). FAM92A1 is a BAR domain protein required for mitochondrial ultrastructure and function. The Journal of Cell Biology, 218(1), 97-111.

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Lipid-Protein Interaction Study Protocol

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Cholesterol (Chol), egg sphingomyelin (SM), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylserine (POPS), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) were obtained from Avanti Polar Lipids, Inc. (Alabaster, AL, USA). The cholesterol analog cholestane spin-label (CSL), bovine eye lens α-crystallin (C4163), HEPES, and sodium chloride (NaCl) were obtained from Sigma Aldrich (St. Louis, MO, USA). Bovine α-crystallin purchased from Sigma Aldrich was used without further purification. The average molecular weight of the α-crystallin subunit was determined to be 20.35 kDa based on the information αA = 19.8 kDa, αB = 22 kDa, and αA:αB = 3:1 from Sigma Aldrich.

Timsina R., Trossi-Torres G., Thieme J., O’Dell M., Khadka N.K, & Mainali L. (2022). Alpha-Crystallin Association with the Model of Human and Animal Eye Lens-Lipid Membranes is Modulated by Surface Hydrophobicity of Membranes. Current eye research, 47(6), 843-853.

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Biophysical Characterization of Antimicrobial Peptide LL-III

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The peptide lasioglossin-III (LL-III) with the sequence NH 2 -VNWKKILGKIIKVVK-CONH 2 was purchased from Primm Srl (Milano, Italy) with a purity of 495%. The lipids cholesterol (Chol), 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC), 1-palmitoyl-2oleoyl-sn-glycero-3-phosphatidylserine (POPS) and 1-palmitoyl-2-oleoyl-glycero-3-phosphatidylcholine (POPC) as well as spin-labeled phosphatidylcholines used as EPR probes (1-palmitoyl-2-stearoyl-(n-doxyl)-sn-glycero-3-phosphatidylcholine, nPC-SL, n = 5, 7, 10, and 14) were provided by Avanti Polar Lipids (Alabaster, United States). The fluorescent probes, 1,6-diphenylhexatriene (DPH) and 6-dodecanoyl-2-dimethylaminonaphthalene (Laurdan), chloroform, methanol, dimethylformamide (DMF), cacodylic acid, calcium chloride (CaCl 2 ), sodium chloride (NaCl), sodium hydroxide (NaOH) and carboxyfluorescein (CF) were acquired from Merck (Darmstadt, Germany). Sodium cacodylate buffer (10 mM) was prepared by solubilizing cacodylic acid in deionized water and adjusting the pH to 6.5 or 7.0 by adding an appropriate amount of NaOH. The pH values of 6.5 and 7.0 were selected to mimic tumor and healthy cell extracellular matrices, respectively. 17

Campanile M., Oliva R., D'Errico G., Del Vecchio P, & Petraccone L. (2023). The anticancer peptide LL-III alters the physico-chemical properties of a model tumor membrane promoting lipid bilayer permeabilization. Physical chemistry chemical physics : PCCP, 25(5).

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Lipid Membrane Characterization Protocol

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Cholesterol (Chol), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC), egg sphingomyelin (SM), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylserine (POPS), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), were purchased from Avanti polar lipids (Alabaster, AL). As per the manufacturer specification, egg SM consists of 86% palmitoyl SM. Chloroform solution of these lipid solutions was used without further purification. ACS grade HEPES, NaCl, and CaCl₂ were purchased from Sigma Aldrich (St. Louis, MO). Buffers were prepared with de-ionized (DI) water. Buffer A (10 mM HEPES, 150 mM NaCl, 5 mM CaCl₂, pH = 7.4) and buffer B (10 mM HEPES, 150 mM NaCl, pH = 7.4) were filtered with a 0.2 μm pore size filter before use.

Felden B., Massire C., Westhof E., Atkins J.F, & Gesteland R.F. (2001). Phylogenetic analysis of tmRNA genes within a bacterial subgroup reveals a specific structural signature. Nucleic Acids Research, 29(7), 1602-1607.

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Membrane Interactions with α-Crystallin

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Lipids, 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC), egg sphingomyelin (SM), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylserine (POPS), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), and cholesterol (Chol), were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL, USA). The bovine eye lens α-crystallin, cholesterol analog cholestane spin-label (CSL), HEPES, and sodium chloride (NaCl) were purchased from Sigma Aldrich (St. Louis, MO, USA). The α-crystallin (C4163) was used without further purification. HEPES buffer (10 mM HEPES, 100 mM NaCl, pH = 7.4) was used to dissolve α-crystallin, prepare membranes, and prepare mixed α-crystallin and membrane samples for EPR measurements. The Sigma Aldrich provided information that αA = 19.8 kDa, αB = 22 kDa, and αA:αB = 3:1. Based on this information, the average molecular weight of the α-crystallin subunit was estimated to be 20.35 kDa.

Timsina R., Trossi-Torres G., O’Dell M., Khadka N.K, & Mainali L. (2021). Cholesterol and Cholesterol Bilayer Domains Inhibit Binding of Alpha-Crystallin to the Membranes Made of the Major Phospholipids of Eye Lens Fiber Cell Plasma Membranes. Experimental eye research, 206, 108544.

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Lipid Composition and Characterization

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Phospholipids 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (PC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylglycerol (PG), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylethanolamine (PE), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylserine (PS), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidic acid (PA), and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(7-nitro-2-1,3-benzoxadiazol-4-yl) (N-NBD-PE) were purchased from Avanti Polar Lipids Inc. (Birmingham, AL, USA). N-dodecyl-β-maltoside (DDM) and n-octyl-β-d-glucoside (OG) were obtained from Glycon (Luckenwalde, Germany). The ionophores valinomycin and CCCP, the pH-sensitive dye ACMA, and all other chemicals and regents were from Sigma-Aldrich (München, Germany), if not stated otherwise. ACMA was dissolved in dimethylsulfoxide; valinomycin in ethanol.

Paweletz L.C., Holtbrügge S.L., Löb M., De Vecchis D., Schäfer L.V., Günther Pomorski T, & Justesen B.H. (2023). Anionic Phospholipids Stimulate the Proton Pumping Activity of the Plant Plasma Membrane P-Type H+-ATPase. International Journal of Molecular Sciences, 24(17), 13106.

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Biophysical Analysis of β-Crystallin Membrane Interactions

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Fresh ~2-year-old bovine lenses were acquired from Pel-Freez Biologicals (Rogers, AZ, USA) and stored at −80° C until use. Cholesterol (Chol), sphingomyelin (SM), and phospholipids (PLs): 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylCholine (POPC), 1-palmditoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE), and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylserine (POPS) were obtained in chloroform from Avanti Polar Lipids, Inc. (Alabaster, AL, USA). Cholesterol analog cholestane spin-label (CSL), HEPES, Tris-HCl, NaN₃, and sodium chloride (NaCl) were obtained from Sigma Aldrich (St. Louis, MO, USA). CSL spin-label was dissolved in chloroform, and the native β_L-crystallin was extracted and purified from the bovine lens cortex and stored in HEPES buffer (10 mM HEPES, 100 mM NaCl, pH = 7.4), as described in Section 4.2. All preparations of β_L-crystallin and the model lens lipid membranes, as well as associated studies into the β_L-crystallin membrane interactions, were performed in HEPES buffer (10 mM HEPES, 100 mM NaCl, pH = 7.4).

Hazen P., Trossi-Torres G., Khadka N.K., Timsina R, & Mainali L. (2023). Binding of βL-Crystallin with Models of Animal and Human Eye Lens-Lipid Membrane. International Journal of Molecular Sciences, 24(17), 13600.

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