title: Structural basis of human erythrocyte glucose transporter function: pH effects on intrinsic fluorescence.
abstract: The effects of pH on the intrinsic fluorescence of purified human erythrocyte glucose transporter (HEGT) were studied to deduce the structure and the ligand-induced dynamics of this protein. D-Glucose increases tryptophan fluorescence of HEGT at a 320-nm peak with a concomitant reduction in a 350-nm peak, suggesting that glucose shifts a tryptophan residue from a polar to a nonpolar environment. Cytochalasin B or forskolin, on the other hand, only produces a reduction at the 350-nm peak. The pH titration of the intrinsic fluorescence of HEGT revealed that at least two tryptophan residues are quenched, one with a pKa of 5.5, the other with a pKa of 8.2, indicating involvement of histidine and cysteine protonation, respectively. D-Glucose abolishes both of these quenchings. Cytochalasin B or forskolin, on the other hand, abolishes the histidine quenching but not the cysteine quenching and induces a new pH quenching with a pKa of about 4, implicating involvement of a carboxyl group. These results, together with the known primary structure and the transmembrane disposition of this protein, predict the dynamic interactions between Trp388 and His337, Trp412 and Cys347, and Trp412 and Glu380, depending on liganded state of HEGT, and suggest the importance of the transmembrane helices 9, 10, and 11 in transport function.
authors.affiliation: Department of Biophysical Sciences, State University of New York, Buffalo.
authors.forename: J J
authors.initials: JJ
authors.lastname: Chin
authors.forename: B H
authors.initials: BH
authors.lastname: Jhun
authors.forename: C Y
authors.initials: CY
authors.lastname: Jung
mesh_terms: D000595:Amino Acid Sequence; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000378:metabolism*; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000378:metabolism
chemical_list: D009004:Monosaccharide Transport Proteins; D014364:Tryptophan