Fractionation of human red cell membrane proteins by ion-exchange chromatography in detergent on Mono Q, with special reference to the glucose transporter.
The anion exchanger Mono Q has been used for rapid and efficient fractionation of human red cell membrane proteins in the easily removable detergents n-octyl-beta-D-glucopyranoside or nonanoyl-N-methylglucamide. In practice the chromatographic resolution of membrane proteins was lower than for water-soluble proteins, perhaps due to protein-protein interactions and microheterogeneity, but several components, or groups of components, separated well upon salt gradient elution. The glucose transporter (or transportase) was eluted early, glycophorins later, and the anion transporter still later. The detergents Berol 185 and the zwitter-ionic derivatives of cholate, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulphonat e and 3-[(3-cholamidopropyl)dimethylammonio]-2-hydroxy-1-propan esulphonate, gave similar chromatographic results but differed in solubilization selectivity. A relatively pure material was also fractionated; viz., a glucose transportase which had been prepared by DEAE-cellulose chromatography. Mono Q, in the presence of octyl glucoside, afforded additional purification, which made automatic sequence determination possible for eighteen amino acid residues. The results indicate that two polypeptides were present in about equimolar amounts.
D000595:Amino Acid Sequence; D000837:Anion Exchange Resins; D001798:Blood Proteins / Q000302:isolation & purification; D002352:Carrier Proteins / Q000097:blood; D002852:Chromatography, Ion Exchange / Q000379:methods; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000032:analysis*; D006801:Humans; D008565:Membrane Proteins / Q000302:isolation & purification*; D009004:Monosaccharide Transport Proteins; D012117:Resins, Synthetic
D000837:Anion Exchange Resins; D001798:Blood Proteins; D002352:Carrier Proteins; D003902:Detergents; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D012117:Resins, Synthetic; C042855:Mono Q