Skip to main content
Log in

Different binding sites for glucose and sorbose at the erythrocyte membrane, studied by gel filtration and infrared spectroscopy

  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

Human red blood cell membranes were solubilized with sodium dodecylsulfate and incubated with various concentrations of14C-glucose and14C-sorbose. After gel filtration on Sephadex G-100, which separated lipoproteins of differing lipid content, it was observed that the radioactivity of the bound glucose coincided with the protein peak. Radioactivity of bound sorbose was found mainly before and after the protein peak. This distribution of bound sugars was confirmed by double labeling experiments in which3H-glucose and14C-sorbose were applied simultaneously. Infrared spectroscopy revealed differences between the membranes loaded with sorbose and glucose. Particularly, the band in the C−O−C and P=O region at 1,225 cm−1 was intensified in the sorbose-loaded membranes. Compared to serum albumin, the erythrocyte membranes were found to bind 4 times as much14C-glucose per mg of protein. It is concluded from the results obtained by gel filtration that glucose and sorbose preferentially bind at different sites of the erythrocyte membrane. The results obtained by infrared spectroscopy correspond with this conclusion.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Bakerman, S., Wasemiller, G. 1967. Studies of the structural units of human erythrocyte membrane. I. Separation, isolation and partial characterization.Biochemistry 6:1100.

    PubMed  Google Scholar 

  • Blumenfeld, O. O. 1968. The proteins of the erythrocyte membrane obtained by solubilization with aqueous pyridine solution.Biochem. Biophys. Res. Commun. 30:200.

    PubMed  Google Scholar 

  • Chapman, D., Kamat, V. B., Levene, R. J. 1968. Infrared spectra and the chain organization of erythrocyte membranes.Science 160:314.

    PubMed  Google Scholar 

  • Dodge, J. T., Mitchell, C., Hanahan, D. J. 1963. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes.Arch. Biochem. Biophys. 100:119.

    PubMed  Google Scholar 

  • Hoogeveen, J. Th., Juliano, R., Coleman, J., Rothstein, A. 1970. Water-soluble proteins of the human red cell membrane.J. Membrane Biol. 3:156.

    Google Scholar 

  • Hummel, J. P., Dreyer, W. J. 1962. Measurement of protein-binding phenomena by gel filtration.Biochim. Biophys. Acta 63:530.

    PubMed  Google Scholar 

  • Lacko, L. 1967. Specificity of sugar carriers in erythrocytes.Nature 213:523.

    PubMed  Google Scholar 

  • Lacko, L., Burger, M. 1961. Common carrier system for sugar transport in human red cells.Nature 191:881.

    PubMed  Google Scholar 

  • LeFevre, P. G. 1954. The evidence for active transport of monosaccharides across the red cell membrane.Symp. Soc. Exp. Biol. No. VIII, Active Transport and Secretion. p. 118.

    Google Scholar 

  • Lenard, J. 1970. Protein and glycolipid components of human erythrocyte membranes.Biochemistry 9:1129.

    PubMed  Google Scholar 

  • Levine, M., Oxender, D. L., Stein, W. D. 1965. The substrate-facilitated transport of the glucose carrier across the human erythrocyte membrane.Biochim. Biophys. Acta 109:151.

    PubMed  Google Scholar 

  • Levine, M., Stein, W. D. 1966. The kinetic parameters of the monosaccharide transfer system of the human erythrocyte.Biochim. Biophys. Acta 127:179.

    PubMed  Google Scholar 

  • Lieb, W. R., Stein, W. D. 1970. Quantitative predictions of a noncarrier model for glucose transport across the human red cell membrane.Biophys. J. 10:585.

    PubMed  Google Scholar 

  • Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J. 1951. Protein measurement with the Folin phenol reagent.J. Biol. Chem. 193:265.

    PubMed  Google Scholar 

  • Maddy, A. H. 1966. The properties of the protein of the plasma membrane of ox erythrocytes.Biochim. Biophys. Acta 117:193.

    PubMed  Google Scholar 

  • Marchesi, S. L., Steers, E., Marchesi, V. T., Tillack, T. W. 1970. Physical and chemical properties of a protein isolated from red cell membranes.Biochemistry 9:50.

    PubMed  Google Scholar 

  • Marchesi, V. T., Steers, E., Jr. 1968. Selective solubilization of a protein component of the red cell membrane.Science 159:203.

    PubMed  Google Scholar 

  • Miyazawa, T., Blout, E. R. 1961. The infrared spectra of polypeptides in various conformations: Amide I and II bands.J. Amer. Chem. Soc. 83:712.

    Google Scholar 

  • Rosenberg, S. A., Guidotti, G. 1969. Fractionation of the protein components of human erythrocyte membranes.J. Biol. Chem. 244:5118.

    PubMed  Google Scholar 

  • Sen, A. K., Widdas, W. F. 1962. Determination of the temperature and pH dependence of glucose transfer across the human erythrocyte membrane measured by glucose exit.J. Physiol. 160:392.

    PubMed  Google Scholar 

  • Stein, W. D. 1969. Transport proteins. Intra-protein interactions across a fluid membrane as a model for biological transport.J. Gen. Physiol. 54:81S.

    Google Scholar 

  • Tipson, R. S., Isbell, H. S. 1962. Infrared absorption spectra in the study of mutarotational equilibria of monosaccharides.J. Phys. Nat. Bur. Stand. 66A:31.

    Google Scholar 

  • Wallach, D. F. H. 1969. Membrane lipids and the conformations of membrane proteins.J. Gen. Physiol. 54:3S.

    Google Scholar 

  • Wallach, D. F. H., Zahler, P. H. 1966. Protein conformations in cellular membranes.Proc. Nat. Acad. Sci. 56:1552.

    PubMed  Google Scholar 

  • Wilbrandt, W. 1960. The sugar transport across the red cell membrane.In: Symposium on Membrane Transport and Metabolism. A. Kleinzeller and A. Kotyk, editors. p. 205. Publishing House, Czechoslovak Academy of Sciences, Prague.

    Google Scholar 

  • Wood, G. C., Cooper, P. F. 1970. The application of gel filtration to the study of protein-binding of small molecules.Chromatogr. Rev. 12:88.

    PubMed  Google Scholar 

  • Zimmer, G., Lacko, L. 1971. Structural change of human red cell membranes in the glucose-preloaded state.F. E. B. S. 12:333.

    Google Scholar 

  • Zöllner, N., Kirsch, K. 1962. Über die quantitative Bestimmung von Lipoiden (Mikromethode) mittels der vielen natürlichen Lipoiden (allen bekannten Plasmalipoiden) gemeinsamen Sulphophosphovanillin-Reaktion.Z. Ges. Exp. Med. 135:545.

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zimmer, G., Lacko, L. & Günther, H. Different binding sites for glucose and sorbose at the erythrocyte membrane, studied by gel filtration and infrared spectroscopy. J. Membrain Biol. 9, 305–318 (1972). https://doi.org/10.1007/BF01868059

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01868059

Keywords

Navigation