Egg phosphatidylcholine pc

Egg Phosphatidylcholine (PC), CAS: 8002–43–5, 1,2-Dipalmitoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DPPG), CAS: 67232–81–9, 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), CAS: 63–89–8, and Cholesterol (Chol), CAS: 57–88–5, were obtained from Sigma-Aldrich Company Ltd. (Poole, UK). 1,2-dioleoyl-sn-glycero-3-phsphoEthanolamine (DOPE), CAS: 4004–05–1, 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), CAS: 144189–73–1, dimethyldioctadecylammonium bromide (DDA), CAS: 3700–67–2, and trehalose 6,6-dibehenate (TDB), CAS: 66758–35–8 were purchased from Avanti Polar Lipids, Inc., (Alabaster, AL), purity >99% (Table I). Ethanol, CAS: 64–17–5, and mEthanol, CAS: 67–56–1, were obtained from Fisher Scientific (Loughborough, UK). TRIS Ultra Pure, CAS: 77–86–1, was obtained from ICN Biomedicals, Inc., (Aurora, Ohio, US). Propofol (2,6-Bis(isopropyl)phenol), CAS: 2078–54–8, and ovalbumin (chicken egg), CAS: 9006–59–1, were obtained from Sigma-Aldrich Company Ltd., (Poole, UK). Ultrafiltration regenerated cellulose membranes (p\n: U2755–10AE) were obtained from Sigma-Aldrich Company Ltd., (Poole, UK) (10 kDa, pore size 0.22 µm), and Biomax polyethersulfone ultrafiltration membrane discs with 300 kDa cutoff, pore size 0.45 µm (p\n: PBMK06210) from Merck Milipore (Darmstadt, Germany).

Alexa Flour 647, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate (DiI) was obtained from Invitrogen. Sephadex G-25 fine resin, egg phosphatidylcholine (PC), cholesteryl oleate, sodium cyanoborohydride (NaCNBH₃), triethylamine (TEA), 3-aminopropyl-triethoxysilan (APTES), ethanolamine (ETA), sodium deoxycholate and HEPES were from Sigma. 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC) was purchased from Avanti Polar Lipids. Cholesterol linked to boron dipyrromethene difluoride at sterol carbon-24 (cholesterol- Bodipy; C-Bodipy) was synthesized as described^{23 (link)}. Cholesteryl-ester- Bodipy (CE-Bodipy) was synthesized by conjugating linoleic acid to C-Bodipy as described²⁴ .

Liposomes were prepared as previously described^{25 (link),26 (link)}. Briefly, Egg phosphatidylcholine (PC) and phosphatidylserine (PS) (Sigma Aldrich) were taken such that 4.5 μmol of total PLs were comprised of 90% Egg PC and 10% brain PS. The lipid mixture was dried under N₂ and dissolved in reconstitution buffer containing 10 mM HEPES/NaOH (hydroxyethyl-1-piperazine ethanesulfonic acid/sodium hydroxide), pH 7.5, 100 mM NaCl, and 1% (w/v) Triton X-100. The detergent (Triton X-100) was slowly removed by Bio beads with 3 changes of 200 mg Bio-Beads SM2 (Bio-Rad) per ml of liposomes and incubated at 4 °C for 16 h with gentle rotation. The protein fraction was reconstituted into liposomes for functional assay and is termed proteoliposomes which were prepared similar to that of liposomes. To the dissolved lipid mixture, 100 µg of recombinant hPLSCR3 was added after the solubilization step and then detergent was slowly removed by Bio-Beads^{19 (link),21 (link),22 (link)}.