title: Endoglycosidase f cleaves the oligosaccharides from the glucose transporter of the human erythrocyte.
abstract: The glucose transporter from human erythrocytes is a heterogeneously glycosylated protein that runs as a very broad band of average apparent Mr 55 000 upon sodium dodecyl sulfate polyacrylamide gel electrophoresis. When the purified preparation of transporter, solubilized in Triton X-100, was treated with endoglycosidase F, much of it ran as a sharp band of Mr 46 000 upon electrophoresis. Moreover, endoglycosidase F released 80% of the radioactivity in a preparation of the transporter labeled in its oligosaccharides with galactose oxidase and tritiated borohydride, and almost none of the remaining radioactivity was located in the Mr 46 000 band. These results suggest that endoglycosidase F can release virtually all of the carbohydrate linked to the transporter polypeptide. A quantitative analysis of the gels was complicated by partial aggregation of polypeptides that occurs due to prolonged incubation in Triton X-100, but at least 65% of the protein in the preparation of purified transporter is the 46 kDa polypeptide. The extracellular domain of the transporter is very resistant to proteolysis; no cleavage occurred upon treatment of intact erythrocytes with seven different proteases at high concentration.
mesh_terms: D002352:Carrier Proteins / Q000097:blood* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000032:analysis*; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006801:Humans; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008970:Molecular Weight; D009004:Monosaccharide Transport Proteins; D017830:Octoxynol; D009844:Oligosaccharides / Q000378:metabolism*; D011092:Polyethylene Glycols
chemical_list: D002352:Carrier Proteins; D009004:Monosaccharide Transport Proteins; D009844:Oligosaccharides; D011092:Polyethylene Glycols; D017830:Octoxynol; D006026:Glycoside Hydrolases; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase