Isolation of glycophorin with deoxycholate |
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Authors: | Jere P. Segrest Thomas M. Wilkinson Lulu Sheng |
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Affiliation: | Department of Pathology, Biochemistry and Microbiology, University of Alabama in Birmingham Medical Center, Birmingham, AL 35294 U.S.A. |
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Abstract: | In a previous communication we reported that human erythrocyte glycophorin prepared by the lithium diiodosalicylate phenol procedure contains approximately 10 mol of lithium diiodosalicylate per mol of glycophorin, and further we showed that this bound lithium diiodosalicylate is difficult to remove by detergents or organic solvents (Romans, A.Y. and Segrest, J.P. (1978) Biochim. Biophys. Acta 511, 297–301). In the present communication we report an alternative purification procedure for glycophorin in which sodium deoxycholate is substituted for lithium diiodosalicylate; the sodium deoxycholate is subsequently removed by gel filtration. Utilizing this procedure, 25–30 mg glycophorin are obtained per gram of lyophilized erythrocyte ghosts. The glycophorin prepared by the sodium deoxycholate procedure, after a single gel filtration step, contains less than 1 mol of sodium deoxycholate per mol glycophorin and is colorless compared with glycophorin prepared by the lithium diiodosalicylate procedure, which has a distinct reddish-brown cast. |
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Keywords: | Glycophorin isolation Sodium deoxycholate extraction (Human erythrocyte) LIS lithium diiodosalicylate SDS sodium dodecyl sulfate |
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