Partial purification of hog kidney sodium-D-glucose cotransport system by affinity chromatography on a phlorizin polymer.
A brush border membrane fraction isolated from hog kidney cortex was solubilized with 0.5% Triton X-100 and subjected to affinity chromatography on a phlorizin polymer. As demonstrated by transport studies with reconstituted proteoliposomes, the polymer adsorbs the sodium-dependent D-glucose transport system. The latter can be eluted from the polymer by 0.5 M D-glucose. The purified fraction contains 0.4% of the membrane protein extract and exhibits a 20--30-fold higher transport activity than the crude membrane extract. Other brush border membrane proteins such as alkaline phosphatase and aminopeptidase M are markedly reduced in the purified fraction. Thus, affinity chromatography on a phlorizin polymer is a suitable tool for the isolation of the sodium-glucose transport system present in brush border membranes.
D000818:Animals; D001692:Biological Transport; D002352:Carrier Proteins / Q000378:metabolism; D002846:Chromatography, Affinity; D005947:Glucose / Q000378:metabolism*; D007672:Kidney Cortex / Q000378:metabolism*; D007700:Kinetics; D008081:Liposomes; D008871:Microvilli / Q000378:metabolism; D008970:Molecular Weight; D009004:Monosaccharide Transport Proteins; D009005:Monosaccharides / Q000378:metabolism; D010695:Phlorhizin; D012964:Sodium / Q000378:metabolism*; D013552:Swine
D002352:Carrier Proteins; D008081:Liposomes; D009004:Monosaccharide Transport Proteins; D009005:Monosaccharides; D012964:Sodium; D010695:Phlorhizin; D005947:Glucose