HPLC and mass spectrometry–grade acetonitrile (ACN), and a bicinchoninic acid assay kit were purchased from Thermo Fisher Scientific. DMSO-d6, D2O, and 15N-labeled hydroxylamine hydrochloride were purchased from Cambridge Isotope Laboratories. DTT, β-mercaptoethanol, and 7-diethylamino-3-(4-maleimidophenyl)-4-methylcoumarin (CPM) were of the highest purity available and obtained from Sigma-Aldrich. The syntheses of the HNO donor, Angeli’s salt (Na2N2O3, AS) and 15N-labeled Angeli’s salt (15N-AS), were performed as previously described (Bonner and Ravid, 1975 (link); Hughes and Cammack, 1999 (link)). The HNO donor, 2-methylsulfonyl benzene N-hydroxy sulfonamide (2-methylsulfonyl Piloty’s acid, 2-MSPA; Toscano et al., 2011 ; Sabbah et al., 2013 (link); Zhu et al., 2015 (link)), the 15N-labeled HNO donor, 2-methylsulfonyl benzene 15N-hydroxy sulfonamide (15N-2-MSPA; Toscano et al., 2011 ; Sabbah et al., 2013 (link); Zhu et al., 2015 (link)), and the donor byproduct, 2-(methylsulfonyl)benzenesulfinic acid (2-MSSA), were gifts from Cardioxyl Pharmaceuticals. Dodecylphosphocholine (DPC) was purchased from Avanti Polar Lipids. Milli Q water was used for all purifications and experiments.
Dodecylphosphocholine dpc
Manufactured by Avanti Polar Lipids
Sourced in United States
Dodecylphosphocholine (DPC) is a non-ionic detergent commonly used in biochemical and biophysical research. It is a water-soluble compound that can solubilize and stabilize membrane proteins, allowing for their structural and functional studies. DPC has a critical micelle concentration of approximately 1.5 mM, making it a suitable choice for various applications involving the manipulation and analysis of membrane-bound proteins.
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Lab products found in correlation
Dmso d6, by Cambridge Isotopes (1 mentions)
Bicinchoninic acid assay kit, by Thermo Fisher Scientific (1 mentions)
Dithiothreitol (dtt), by Merck Group (1 mentions)
β mercaptoethanol, by Merck Group (1 mentions)
7 diethylamino 3 4 maleimidophenyl 4 methylcoumarin cpm, by Merck Group (1 mentions)
J 715 spectropolarimeter, by Jasco (1 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (1 mentions)
1 1 1 3 3 3 hexafluoro 2 propanol hfip, by Merck Group (1 mentions)
N 7 nitrobenz 2 oxa 1 3 diazol 4 yl 1 2 dihexadecanoyl sn glycero 3 phosphoethanolamine nbd pe, by Thermo Fisher Scientific (1 mentions)
Star red kk114, by Abberior (1 mentions)
3 protocols using dodecylphosphocholine dpc
1
Synthesis and Characterization of HNO Donors
2
CD Spectroscopy for Secondary Structure
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CD measurements for secondary structure estimation were performed on a Jasco J 715 Spectropolarimeter (Jasco, Gross-Umstadt, Germany) at room temperature using quartz cuvettes with an optical path length of 0.02 cm. The CD spectra were measured between 260 nm and 180 nm with a 0.2 nm step resolution. To improve accuracy, 3 scans were accumulated and averaged. The peptides were dissolved in 20 mM phosphate-buffered saline (PBS) (20 mM NaPi, without NaCl, pH 7.4). Spectra were measured in the absence and presence of 20 mM POPS, 20 mM POPC and 0.5 µM Dodecylphosphocholine (DPC) (Avanti Polar Lipids, Alabaster, USA) and 0.5 µM sodium dodecyl sulfate (SDS) (Sigma Life Science, Darmstadt, Germany), with POPS and SDS mimicking cancer, POPC and DPC healthy mammalian membranes, respectively. Liposomes were prepared as described above. The respective peptide to surfactant molar ratios were 25:1. The background signals were subtracted after the measurements. Percentage secondary structure calculations were conducted using Spectra Secondary structure estimation [78 (link)].
3
Peptide Synthesis and Lipid Preparation
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The peptide sequences derived from the gp41 MPER-TMD region displayed in Figure 1b were produced as C-terminal carboxamides by solid-phase synthesis using Fmoc chemistry and purified by HPLC. PC-based synthetic phospholipids and dodecylphosphocholine (DPC) were purchased from Avanti Polar Lipids (Birmingham, AL, USA). N-(7-Nitrobenz-2-Oxa-1,3-Diazol-4-yl)-1,2-Dihexadecanoyl-sn-Glycero-3-Phosphoethanolamine (NBD-PE) was from Thermo Fisher Scientific (Waltham, Massachusetts, USA). Abberior STAR RED (KK114) was obtained from Abberior (Göttingen, Germany). 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP) was obtained from Sigma-Aldrich (St. Louis, Missouri, USA).
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