Interaction of nonionic detergents with phospholipids in hepatic microsomes at subsolubilizing concentrations as studied by 31P-NMR |
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| Affiliation: | 1. University of Groningen, University Medical Center Groningen, Department of Genetics, Groningen, The Netherlands;2. Netherlands Heart Institute, Utrecht, The Netherlands;3. University of Groningen, University Medical Center Groningen, Department of Clinical and Experimental Cardiology, Groningen, The Netherlands;4. Cardiology Clinical Academic Group, St. George''s University of London, Cranmer Terrace, London, United Kingdom;5. Departments of Cardiac, Thoracic, and Vascular Sciences, University of Padua, Padua, Italy;6. Department of Clinical Genetics, Amsterdam Cardiovascular Sciences, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands;7. Durrer Center for Cardiovascular Research, Netherlands Heart Institute, Utrecht, The Netherlands;8. Department of Pathology, Beth Israel Deaconess Medical Center & Harvard Medical School, Boston, MA, USA;9. University of Groningen, University Medical Center Groningen, Department of Pathology, Groningen, The Netherlands;1. Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children''s Hospital, Boston, MA, USA;2. Center for Human Genetics, University Hospitals Cleveland Medical Center, Cleveland, OH, USA;3. Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, OH, USA;4. Center for Inherited Disorders of Energy Metabolism (CIDEM), University Hospitals Cleveland Medical Center, Cleveland, OH, USA;5. Department of Pediatrics, Harvard Medical School, Boston, MA, USA;6. Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA;7. Department of Pharmacology, Case Western Reserve University, Cleveland, OH, USA;8. Department of Medicine, Case Western Reserve University, Cleveland, OH, USA;9. Department of Neurology, Boston Children''s Hospital, Boston, MA, USA;10. Department of Pediatrics, Case Western Reserve University, Cleveland, OH, USA |
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| Abstract: | The effect of low concentrations of nonionic detergents with different critical micelle concentrations such as Triton X-100, Brij 35 and octylglucoside on rabbit liver microsomes is studied by means of 31P-NMR, 1H-NMR, dynamic light scattering and functional investigations. Hexane phosphonic acid diethyl ester was used as a phosphorus membrane probe molecule to monitor the interaction of detergent molecules with microsomal phospholipids by 31P-NMR. This method is more sensitive than 31P-NMR of phospholipids alone and permitted the estimation of the maximum number of detergent molecules which can be incorporated in microsomes without the formation of mixed micelles outside the membrane. These membrane saturation concentrations were determined to be 0.07 (Brij 35), 0.1 (Triton X-100) and 0.4 (octylglucoside) (molar ratio of detergent/total phospholipids). Above these detergent concentrations, mixed micelles consisting of detergent and membrane constituents are formed, coexisting with the microsomes up to the membrane solubilization concentration. The results indicate a dependence of the membrane saturation concentration on the critical micelle concentration of the detergent and a preferential removal of phosphatidylcholine over phosphatidylethanolamine from the microsomes by all detergents studied. |
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