High-performance liquid chromatographic separation of membrane proteins isolated from erythrocyte ghosts

Membrane proteins extracted from erythrocyte ghosts with sodium dodecyl sulphate (SDS), 3-(3-cholamidopropyl)-dimethylamminopropane sulfonate (CHAPS) or octylglucoside have been analyzed in various reversed-phase high-performance liquid chromatographic systems. Only SDS was able to solubilize consid...

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Bibliographic Details
Published in:Journal of Chromatography A 1989, Vol.462, p.255-268
Main Authors: Welinder, Benny S., Sørensen, Hans H., Hansen, Bruno
Format: Article
Language:English
Subjects:
Analytical, structural and metabolic biochemistry
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
General aspects, investigation methods
Proteins
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Summary:Membrane proteins extracted from erythrocyte ghosts with sodium dodecyl sulphate (SDS), 3-(3-cholamidopropyl)-dimethylamminopropane sulfonate (CHAPS) or octylglucoside have been analyzed in various reversed-phase high-performance liquid chromatographic systems. Only SDS was able to solubilize considerable amounts of membrane proteins with mol.wt. > 15 000 daltons, but these membrane proteins were recovered in poor yield from a silica-based C 4 column eluted with an acetonitrile gradient in trifluoroacetic acid (TFA). A resin-based phenyl column eluted with a similar TFA-acetonitrile gradient was found to be a better choice with respect to the recovery of membrane proteins with mol.wt. > 15 000 daltons, and when this column was eluted with an acetic acid gradient with increasing amounts of acetonitrile, erythrocyte ghost membrane proteins solubilized in SDS (mol.wt. 10 000–200 000 daltons) were separated in six major and several minor components with satisfactory recovery.
ISSN:0021-9673
DOI:10.1016/S0021-9673(00)91352-9