Use of mild detergent gel electrophoresis for isolation and characterization of the kidney brush border D-glucose transporter.
Gradient gel electrophoresis was performed under mild detergent conditions to separate pig kidney brush border membrane proteins and to identify the smallest functional molecular protein entity of the D-glucose transporter. The various protein bands obtained from the nondenaturing gel system in a semipreparative scale were eluted by electrodialysis. These proteins were then reintegrated into proteoliposomes and tested for D-glucose-inhibitable [3H]phlorizin binding. The D-glucose transporter had a molecular mass of 70 kDa in mild detergent electrophoresis conditions and in subsequent SDS analysis.
Laboratoire de Biochimie, Faculté de Médecine, Nice, France.
M
M
Starita-Geribaldi
J C
JC
Poiree
P
P
Sudaka
D000818:Animals; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D007672:Kidney Cortex / Q000378:metabolism*; D008081:Liposomes; D008871:Microvilli / Q000378:metabolism*; D008970:Molecular Weight; D009004:Monosaccharide Transport Proteins / Q000302:isolation & purification* / Q000378:metabolism; D017830:Octoxynol; D011092:Polyethylene Glycols; D011510:Proteolipids; D013552:Swine
D003902:Detergents; D008081:Liposomes; D009004:Monosaccharide Transport Proteins; D011510:Proteolipids; C020485:proteoliposomes; D011092:Polyethylene Glycols; D017830:Octoxynol