Summary
We have previously shown that the human red cell glucose transport protein and the anion exchange protein, band 3, are in close enough contact that information can be transmitted from the glucose transport protein to band 3. The present experiments were designed to show whether information could be transferred in the reverse direction, using changes in tryptophan fluorescence to report on the conformation of the glucose transport protein. To see whether tryptophan fluorescence changes could be attributed to the glucose transport protein, we based our experiments on procedures used by Helgerson and Carruthers [Helgerson, A.L., Carruthers, A., (1987)J. Biol. Chem. 262:5464–5475] to displace cytochalasin B (CB), the specificd-glucose transport inhibitor, from its binding site on the inside face of the glucose transport protein, and we showed that these procedures modified tryptophan fluorescence. Addition of 75mm maltose, a nontransportable disaccharide which also displaces CB, caused a timedependent biphasic enhancement of tryptophan fluorescence in fresh red cells, which was modulated by the specific anion exchange inhibitor, DBDS (4,4′-dibenzamido-2,2′-stilbene disulfonate). In a study of nine additional disaccharides, we found that both biphasic kinetics and DBDS effects depended upon specific disaccharide conformation, indicating that these two effects could be attributed to a site sensitive to sugar conformation. Long term (800 sec) experiments revealed that maltose binding (±DBDS) caused a sustained damped anharmonic oscillation extending over the entire 800 sec observation period. Mathematical analysis of the temperature dependence of these oscillations showed that 2 μm DBDS increased the damping term activation energy, 9.5±2.8 kcal mol−1 deg−1, by a factor of four to 39.7±5.1 kcal mol−1 deg−1, providing strong support for the view that signalling between the glucose transport protein and band 3 goes in both directions.
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References
Alvarez, J., Lee, D.C., Baldwin, S.A., Chapman, C. 1987. Fourier transform infrared spectroscopic study of the structure and conformational changes in the human erythrocyte glucose transporter.J. Biol. Chem. 262:3502–3509
Appleman, J.R., Lienhard, G.E. 1985. Rapid kinetics of the glucose transporter from human erythrocytes.J. Biol. Chem. 260:4575–4578
Barnett, J.E.G., Holman, G.D., Chalkley, R.A., Munday, K.A. 1975. Evidence for two asymmetrical conformational states in the human erythrocyte sugar-transport system.Biochem. J. 145:417–429
Barnett, J.E.G., Holman, G.D., Munday, K.A. 1973. An explanation of the asymmetric binding of sugars to the human erythrocyte sugar-transport systems.Biochem. J. 135:539–541
Bennett, V., Stenbuck, P.J. 1980. Association between ankyrin and the cytoplasmic domain of band 3 isolated from the human erythrocyte membrane.J. Biol. Chem. 255:6424–6432
Carruthers, A. 1986. Anomalous asymmetric kinetics of human red cell hexose transfer: Role of cytosolic adenosine 5′-triphosphate.Biochemistry 25:3592–3602
Chance, B., Pye, E.K., Ghosh, A.K., Hess, B. (editors) 1973. Biological and Biochemical Oscillators. Academic, New York
Dix, J.A., Verkman, A.S., Solomon, A.K., Cantley, L.C. 1979. Human erythrocyte anion exchange site characterized using a fluorescent probe.Nature 282:520–522
Duhm, V.J., Deuticke, B., Gerlach, E. 1969. Metabolism of 2,3-diphosphoglycerate and glycolysis in human erythrocytes. The influence of sulfate, tetrathionate and disulfite.Hoppe-Seyler's Z. Physiol. Chem. 350:1008–1016
Fossel, E.T., Solomon, A.K. 1978. Ouabain-sensitive interaction between human red cell membrane and glycolytic enzyme complex in cytosol.Biochim. Biophys. Acta 510:99–111
Gorga, F.R., Lienhard, G.E. 1982. Changes in the intrinsic fluorescence of the human erythrocyte monosaccharide transporter upon ligand binding.Biochemistry 21:1905–1908
Helgerson, A.L., Carruthers, A. 1987. Equilibrium ligand binding to the human erythrocyte sugar transporter.J. Biol. Chem. 262:5464–5475
Janoshazi, A., Solomon, A.K. 1989. Interaction among anion, cation and glucose transport proteins in the human red cell.J. Membrane Biol. 112:25–37
Janoshazi, A., Solomon, A.K. 1990. Interaction between red cell glucose transport protein and band 3.Biophys. J. 57:96a
Kahlenberg, A., Dolansky, D. 1972. Structural requirements ofd-glucose for its binding to isolated human erythrocyte membranes.Can. J. Biochem. 50:638–643
Kotaki, A., Naoi, M., Yagi, K. 1971. A diaminostilbene dye as a hydrophobic probe for proteins.Biochim. Biophys. Acta 224:547–556
Krupka, R.M. 1971. Evidence for a carrier conformational change associated with sugar transport in erythrocytes.Biochemistry 10:1143–1148
LeFevre, P.G. 1948. Evidence of active transfer of certain non-electrolytes across the human red cell membrane.J. Gen. Physiol. 31:505–527
Lew, V.L. 1971. On the ATP dependence of the Ca2+-induced increase in K+ permeability observed in human red cells.Biochim. Biophys. Acta 233:827–830
May, J.M. 1987. Labelling of human erythrocyte band 3 with maltosylisothiocyanate.J. Biol. Chem. 262:3140–3145
Miller, D.M., Olson, J.S., Pflugrath, J.W., Quiocho, F.A. 1983. Rates of ligand binding to periplasmic proteins involved in bacterial transport and chemotaxis.J. Biol. Chem. 258:13665–13672
Mueckler, M. 1989. Structure and function of the glucose transporter.In: Red Blood Cell Membranes—Structure—Function—Clinical Implications. P. Agre and J.C. Parker, editors. pp: 31–45. Marcel Dekker, New York
Mueckler, M., Caruso, C., Baldwin, S.A., Panico, M., Blench, I., Morris, H.R., Allard, W.J., Lienhard, G.E., Lodish, H.F. 1985. Sequence and structure of a human glucose transporter.Science 229:941–945
Mullins, R.E., Langdon, R.G. 1980a. Maltosyl isothiocyanate: An affinity label for the glucose transporter of the human erythrocyte membrane. 1. Inhibition of glucose transport.Biochemistry 19:1199–1205
Mullins, R.E., Langdon, R.G. 1980b. Maltosyl isothiocyanate: An affinity label for the glucose transporter of the human erythrocyte membrane. 2. Identification of the transporter.Biochemistry 19:1205–1212
Pimentel, G.C., McClellan, A.L. 1960. The Hydrogen Bond. Table 7-II. Freeman, San Francisco
Quiocho, F.A. 1986. Carbohydrate-binding proteins: Tertiary structures and protein-sugar interactions.Annu. Rev. Biochem. 55:287–315
Quiocho, F.A. 1989. Protein-carbohydrate interactions: Basic molecular features.Pure Appl. Chem. 61:1293–1306
Rees, W.D., Gliemann, J., Holman, G.D. 1987. Re-examination of hexose-transporter inhibition and labelling by hexose isothiocyanates.Biochem. J. 241:857–862
Rogalski, A.A., Steck, T.L., Waseem, A. 1989. Association of glyceraldehyde-3-phosphate dehydrogenase with the plasma membrane of the intact human red blood cell.J. Biol. Chem. 264:6438–6446
Strapazon, E., Steck, T.L. 1977. Interaction of aldolase and the membrane of human erythrocytes.Biochemistry 16:2966–2971
Verkman, A.S., Dix, J.A., Solomon, A.K. 1983. Anion transport inhibitor binding to band 3 in red blood cell membranes.J. Gen. Physiol. 81:421–449
Verkman, A.S., Lukacovic, M.F., Tinklepaugh, M.S., Dix, J.A. 1986. Quenching of red cell tryptophan fluorescence by mercurial compounds.Membrane Biochem. 6:269–289
Vyas, N.K., Vyas, M.N., Quiocho, F.A. 1988. Sugar and signaltransducer binding sites of theEscherichia coli galactose chemoreceptor protein.Science 242:1290–1295
Walmsley, A.R. 1988. The dynamics of the glucose transporter.Trends Biochem. Sci. 13:226–231
Widdas, W.F. 1988. Old and new concepts of the membrane transport for glucose in cells.Biochim. Biophys. Acta 947:385–404
Yu, J., Steck, T.L. 1975. Associations of band 3, the predominant polypeptide of the human erythrocyte membrane.J. Biol. Chem. 250:9176–9184
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Janoshazi, A., Kifor, G. & Solomon, A.K. Conformational changes in human red cell membrane proteins induced by sugar binding. J. Membrain Biol. 123, 191–207 (1991). https://doi.org/10.1007/BF01870403
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DOI: https://doi.org/10.1007/BF01870403