1 2 dimyristoyl sn glycero 3 phosphocholine

1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) was purchased from Lipoid GmbH (>99%, Ludwigshafen, Germany). The pure saponin aescin (≥95%, CAS number 6805-41-0), cholesterol (≥95%), and chloroform were obtained from Sigma Aldrich (Munich, Germany). Samples for DSC, SAXS and WAXS were prepared with deionized, purified water (Sartorius arium VF pro, Göttingen, Germany). SANS samples were prepared in deuterium oxide (>99%, Carl Roth GmbH + Co. KG, Karlsruhe, Germany). Since aescin dissolves poorly in water and decreases the pH significantly, a 50 mM phosphate buffer (pH 7.4/pD 7.8) was used to guarantee a constant pH and solubility in aqueous solution. DMPC was used at a mass concentration of 15 g · L⁻¹. The aescin concentration used (in mol%) is calculated by taking into account all components (DMPC and cholesterol) in solution (DMPC, aescin, and cholesterol): $x(aescin)=n_{aescin}/(n_{aescin}+n_{DMPC}+n_{cholesterol})$ . The cholesterol concentration is defined in dependence of the amount of DMPC: $x(cholesterol)=n_{cholesterol}/(n_{DMPC}+n_{cholesterol})$ .

DMU-214 was synthesized as described elsewhere (Androutsopoulos et al., 2011 (link)). The identity and purity of the compound were confirmed using NMR and LC–MS. 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC, 14:0 PC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC, 16:0 PC) were kindly provided by Lipoid GmbH Ludwigshafen am Rhein, Germany. 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC, 16:0-18:1 PC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) sodium salt (POPG) were purchased from Avanti Polar Lipids Inc. (Alabaster, AL, USA).

1,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC) was a kind gift from Lipoid (Ludwigshafen, Germany). 1-Palmitoyl-2-stearoyl-(5-doxyl)-sn-glycero-3-phosphocholine (16:0-5 doxyl PC), 1-palmitoyl-2-stearoyl-(12-doxyl)-sn-glycero-3-phosphocholine (16:0-12 doxyl PC), 1-palmitoyl-2-stearoyl-(16-doxyl)-sn-glycero-3-phosphocholine (16:0-16 doxyl PC) were bought from Sigma-Aldrich (St. Louis, MO, USA). DIBMA copolymer, commercially available under the trade name PureCube DIBMA 10 in TRIS (lyophilized in Tris-buffer, pH 7.5), was purchased from Cube Biotech (Monheim am Rhein, Germany). N-Dodecyl-ß-D-maltoside (DDM) was acquired from Anatrace (Affymetrix, Cleveland, OH, USA). Isopropyl-β-D-thiogalactopyranosid (IPTG) was bought from Carl Roth (Karlsruhe, Germany). EDTA-free complete protease inhibitor cocktail was procured from Roche Life Science (Mannheim, Germany). Ni-NTA superflow agarose was purchased from Qiagen (Hilden, Germany). All other reagents were of analytical grade.
Protein concentrations were determined using an UV-VIS spectrophotometer (UV-2450, Shimadzu Corporation, Kyoto, Japan). The concentration of NpSRII was determined by using the known extinction coefficient of 50,000 M⁻¹ cm⁻¹ at λ = 280 nm

Palladium tetraphenyltetrabenzoporphyrin (1) was purchased from Frontier Scientific, Inc. (Logan, UT, USA). Perylene (2) was purchased from Sigma-Aldrich Chemie BV (Zwijndrecht, The Netherlands). The synthesis of 2,5,8,11-tetra(tert-butyl)perylene (compound 3) is described elsewhere [25 (link)]. Sodium N-(carbonyl-methoxypolyethylene glycol-2000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE-mPEG-2000), 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) were purchased from Lipoid GmbH (Ludwigshafen, Germany) and stored at −18 °C. Dulbecco’s phosphate buffered saline (PBS) was purchased from Sigma Aldrich and had a formulation of 8 g/L NaCl, 0.2 g/L KCl, 0.2 g/L KH₂PO₄, and 1.15 g/L K₂HPO₄ with a pH of 7.1–7.5. Antioxidant supplemented PBS was prepared by dissolving l-ascorbic acid and/or glutathione in PBS and neutralizing with sodium hydroxide to pH 7.0–7.6. Other chemicals were purchased from major chemical suppliers and used as received. Data was processed with Origin Pro and Microsoft Excel software. Photodissociation experiments using blue light were recorded in a Varian Cary 50 spectrometer (Agilent Technologies, Amstelveen, The Netherlands) equipped with a Cary Single Cell Peltier for temperature control.

Highly pure phospholipids were used. Egg phosphatidylcholine, (EPC S, 99.1% pure, T_m -5°C, batch no. 108064-03/103); hydrogenated soybean phosphatidylcholine (HSPC, ≥98% pure, T_m 52.5°C, batch no. 256270-1/58); 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC, ≥99 pure, T_m 24°C, batch no. 562212-1/25); 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC, ≥99 pure, T_m 41.4°C, batch no. 563098-1/905); and cholesterol (NF grade) were all obtained from Lipoid (Ludwigshafen, Germany). PEG-DSPE-2000 was obtained from Genzyme Pharmaceuticals (Liestal, Switzerland).

Synthetic phospholipids 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPM) were purchased from Lipoid GmbH (Ludwigshafen, Germany). The other lipid components of liposomes, Cholesterol (≥99%), α-Tocopherol acetate (≥96%), and Ubiquinol (USP reference standard), were supplied by Sigma Aldrich, as well as the aqueous compounds Ribitol (Adonitol, ≥99%), Taurine (≥99%), Sodium tetraborate (Na₂B₄O₇, ≥99.5%), and Boric Acid (H₃BO₃, ≥99.5%). Brimonidine was obtained in Fagron Iberica (Terrassa, Spain) and travoprost in MedChemExpress (Monmouth Junction, NJ, USA).
Defined Trypsin Inhibitor was supplied by Life Technologies (Madrid, Spain). Fetal Bovine Serum was supplied by Thermo Fisher (Madrid, Spain). Trypan-Blue, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide], Dimethyl sulfoxide (DMSO), Dulbecco’s phosphate-buffered saline (DPBS), Trypsin-EDTA 0.05%, Sodium Chloride (NaCl), Sodium Chloride 5M, and 2% Gelatin solution were provided by Sigma Aldrich (Madrid, Spain). T-75 flasks and polystyrene 15 mL tubes were from Sarstedt (Madrid, Spain).