Evidence from deuterium nuclear magnetic resonance for the temperature-dependent reversible self-association of erythrocyte band 3 in dimyristoylphosphatidylcholine bilayers.
Band 3, isolated from human erythrocytes, has been reconstituted into bilayers of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) deuterated in the terminal methyl groups of the choline head group. By use of Triton X-100 for selective extraction and purification of band 3 and then cholate for sub...
| Main Authors: | , , |
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| 格式: | Journal article |
| 語言: | English |
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1986
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| _version_ | 1826287142865731584 |
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| author | Dempsey, C Ryba, N Watts, A |
| author_facet | Dempsey, C Ryba, N Watts, A |
| author_sort | Dempsey, C |
| collection | OXFORD |
| description | Band 3, isolated from human erythrocytes, has been reconstituted into bilayers of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) deuterated in the terminal methyl groups of the choline head group. By use of Triton X-100 for selective extraction and purification of band 3 and then cholate for subsequent solubilization with the lipid, a number of reconstituted complexes were produced by exhaustive detergent dialysis with protein:lipid weight ratios of between 0.32:1 and 1.25:1. Electron micrographs of negatively stained complexes showed that this method produced large vesicles of greater than 300-nm diameter. Deuterium nuclear magnetic resonance (NMR) spectra from the choline methyl deuterons in bilayer lipid above the liquid-crystal-gel phase transition temperature were shown to change systematically with increasing concentrations of band 3 in the bilayers. The measured quadrupole splittings, taken as the separation of the turning points in the recorded spectra, decreased from a value of 1.28 kHz for pure lipid to 0.98 kHz for bilayers with a protein:lipid ratio of 1.25:1 at 26 degrees C. At 35 degrees C, a more pronounced decrease in the quadrupole splittings was measured. The data from the complexes with protein:lipid ratios up to 0.7:1 (w/w) obey the mathematical treatment for a rapid two-site exchange between lipids at the protein-lipid interface and the bulk lipid phase. The temperature dependence of the measured quadrupole splitting with respect to the protein:lipid ratio indicates that the amount of lipid at the protein-lipid interface increases with increasing temperature.(ABSTRACT TRUNCATED AT 250 WORDS) |
| first_indexed | 2024-03-07T01:54:12Z |
| format | Journal article |
| id | oxford-uuid:9b285529-6a00-4bb7-ac53-9c33b8770621 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T01:54:12Z |
| publishDate | 1986 |
| record_format | dspace |
| spelling | oxford-uuid:9b285529-6a00-4bb7-ac53-9c33b87706212022-03-27T00:26:42ZEvidence from deuterium nuclear magnetic resonance for the temperature-dependent reversible self-association of erythrocyte band 3 in dimyristoylphosphatidylcholine bilayers.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:9b285529-6a00-4bb7-ac53-9c33b8770621EnglishSymplectic Elements at Oxford1986Dempsey, CRyba, NWatts, ABand 3, isolated from human erythrocytes, has been reconstituted into bilayers of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) deuterated in the terminal methyl groups of the choline head group. By use of Triton X-100 for selective extraction and purification of band 3 and then cholate for subsequent solubilization with the lipid, a number of reconstituted complexes were produced by exhaustive detergent dialysis with protein:lipid weight ratios of between 0.32:1 and 1.25:1. Electron micrographs of negatively stained complexes showed that this method produced large vesicles of greater than 300-nm diameter. Deuterium nuclear magnetic resonance (NMR) spectra from the choline methyl deuterons in bilayer lipid above the liquid-crystal-gel phase transition temperature were shown to change systematically with increasing concentrations of band 3 in the bilayers. The measured quadrupole splittings, taken as the separation of the turning points in the recorded spectra, decreased from a value of 1.28 kHz for pure lipid to 0.98 kHz for bilayers with a protein:lipid ratio of 1.25:1 at 26 degrees C. At 35 degrees C, a more pronounced decrease in the quadrupole splittings was measured. The data from the complexes with protein:lipid ratios up to 0.7:1 (w/w) obey the mathematical treatment for a rapid two-site exchange between lipids at the protein-lipid interface and the bulk lipid phase. The temperature dependence of the measured quadrupole splitting with respect to the protein:lipid ratio indicates that the amount of lipid at the protein-lipid interface increases with increasing temperature.(ABSTRACT TRUNCATED AT 250 WORDS) |
| spellingShingle | Dempsey, C Ryba, N Watts, A Evidence from deuterium nuclear magnetic resonance for the temperature-dependent reversible self-association of erythrocyte band 3 in dimyristoylphosphatidylcholine bilayers. |
| title | Evidence from deuterium nuclear magnetic resonance for the temperature-dependent reversible self-association of erythrocyte band 3 in dimyristoylphosphatidylcholine bilayers. |
| title_full | Evidence from deuterium nuclear magnetic resonance for the temperature-dependent reversible self-association of erythrocyte band 3 in dimyristoylphosphatidylcholine bilayers. |
| title_fullStr | Evidence from deuterium nuclear magnetic resonance for the temperature-dependent reversible self-association of erythrocyte band 3 in dimyristoylphosphatidylcholine bilayers. |
| title_full_unstemmed | Evidence from deuterium nuclear magnetic resonance for the temperature-dependent reversible self-association of erythrocyte band 3 in dimyristoylphosphatidylcholine bilayers. |
| title_short | Evidence from deuterium nuclear magnetic resonance for the temperature-dependent reversible self-association of erythrocyte band 3 in dimyristoylphosphatidylcholine bilayers. |
| title_sort | evidence from deuterium nuclear magnetic resonance for the temperature dependent reversible self association of erythrocyte band 3 in dimyristoylphosphatidylcholine bilayers |
| work_keys_str_mv | AT dempseyc evidencefromdeuteriumnuclearmagneticresonanceforthetemperaturedependentreversibleselfassociationoferythrocyteband3indimyristoylphosphatidylcholinebilayers AT ryban evidencefromdeuteriumnuclearmagneticresonanceforthetemperaturedependentreversibleselfassociationoferythrocyteband3indimyristoylphosphatidylcholinebilayers AT wattsa evidencefromdeuteriumnuclearmagneticresonanceforthetemperaturedependentreversibleselfassociationoferythrocyteband3indimyristoylphosphatidylcholinebilayers |