Topography and functions of sulfhydryl groups of the human erythrocyte glucose transport mechanism.
Membrane-impermeant and -permeant maleimides were applied to characterize the location and function of the sulfhydryl (SH) groups essential for the facilitated diffusion mediated by the human erythrocyte glucose transport protein. Three such classes have been identified. Type I SH is accessible to membrane-impermeant reagents at the outer (exofacial) surface of the intact erythrocyte. Alkylation of this class inhibits glucose transport; D-glucose and cytochalasin B protect against the alkylation. Type II SH is located at the inner (endofacial) surface of the membrane and is accessible to the membrane-impermeant reagent glutathione maleimide only after lysis of the erythrocyte. D-glucose enhances, while cytochalasin B reduces, the alkylation of Type II SH by maleimides. Reaction of Types I and II SH with an impermeant maleimide increases the half-saturation concentration for binding of D-glucose to erythrocyte membranes. By contrast, inactivation of Type III SH markedly decreases the half-saturation concentration for the binding of D-glucose and other transported sugars. Type III SH is inactivated by the relatively lipid-soluble reagents N-ethylmaleimide (NEM) and dipyridyl disulfide, but not by the impermeant glutathione maleimide. Type III SH is thus located in a hydrophobic membrane domain. A kinetic model constructed to explain these observations indicates that Type III SH is required for the translocation event in a hydrophobic membrane domain which leads to the dissociation of glucose bound to transport sites at the membrane surfaces.
Abbott
R E
RE
Department of Physiology and Cellular Biophysics, Columbia University College of Physicians & Surgeons, New York City, New York.
Schachter
D
D
D000478:Alkylation; D001692:Biological Transport; D001786:Blood Glucose / Q000378:metabolism*; D004058:Diffusion; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D008301:Maleimides / Q000494:pharmacology; D008954:Models, Biological; D013438:Sulfhydryl Compounds / Q000378:metabolism*
D001786:Blood Glucose; D008301:Maleimides; D013438:Sulfhydryl Compounds