Lipids that determine detergent resistance of MDCK cell membrane fractions

• Published in: Chemistry and physics of lipids Vol. 191; pp. 68 - 74
• Main Authors: Manni, Marco M., Cano, Ainara, Alonso, Cristina, Goñi, Félix M.
• Format: Journal Article
• Language: English
• Published: Ireland Elsevier Ireland Ltd 01.10.2015
• Subjects: Animals; Chromatography, High Pressure Liquid; Detergents - chemistry; Detergents - metabolism; Diacylglycerols; Dogs; DRM; Lipid chain-length; Lipid rafts; Lipid unsaturation; Lipidomics; Madin Darby Canine Kidney Cells; Membrane Microdomains - metabolism; Phosphatidylcholines - analysis; Phosphatidylethanolamines - analysis; Phospholipids; Phospholipids - analysis; Principal Component Analysis; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Sphingolipids; Sphingolipids - analysis; Lipidomics; Lipid rafts; Sphingolipids; Phospholipids; DAG; PCA; Cer; PC; Lipid chain-length; UHPLCToF-MS; PE; PI; SM; Diacylglycerols; MDCK; TAG; ChoE; Lipid unsaturation; DRM; CMH
• ISSN: 0009-3084; 1873-2941
• DOI: 10.1016/j.chemphyslip.2015.08.011

•A lipidomic study of mammalian detergent-resistant membranes (DRM) from MDCK cells has been performed.•DRM are enriched in sphingomyelins, ceramides and diacylglycerols with respect to the total cell lipids.•DRM are enriched in saturated over unsaturated acyl chains, and in sn-1 acyl chains containing 16 carbon atoms.•Phosphatidylethanolamines and phosphatidylinositols, that were mainly unsaturated, did not show a preference for DRM. A comparative lipidomic study has been performed of whole Madin-Darby canine kidney epithelial cells and of the detergent-resistant membrane fraction (DRM) obtained after treating the cells with the non-ionic detergent Triton X-100. The DRM were isolated following a standard procedure that is extensively used in cell biology studies. Significant differences were found in the lipid composition of the whole cells and of DRM. The latter were enriched in all the analyzed sphingolipid classes: sphingomyelins, ceramides and hexosylceramides. Diacylglycerols were also preferentially found in DRM. The detergent-resistant fraction was also enriched in saturated over unsaturated fatty acyl chains, and in sn-1 acyl chains containing 16 carbon atoms, over the longer and shorter ones. The glycerophospholipid species phosphatidylethanolamines and phosphatidylinositols, that were mainly unsaturated, did not show a preference for DRM. Phosphatidylcholines were an intermediate case: the saturated, but not the unsaturated species were found preferentially in DRM. The question remains on whether these DRM, recovered from detergent-membrane mixtures by floatation over a sucrose gradient, really correspond to membrane domains existing in the cell membrane prior to detergent treatment.