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PMID: 3223921 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Proteolytic dissection as a probe of conformational changes in the human erythrocyte glucose transport protein.

The Biochemical journal ·Vol. 256 ·No. 2 ·1988-12-01 ·Pages 421-7

Gibbs AF, Chapman D, Baldwin SA

Abstract

Tryptic digestion has been used to investigate the conformational changes associated with substrate translocation by the human erythrocyte glucose transporter. The effects of substrates and inhibitors of transport on the rates of tryptic cleavage at the cytoplasmic surface of the membrane have confirmed previous observations that this protein can adopt at least two conformations. In the presence of phloretin or 4,6-O-ethylidene-D-glucose, the rate of cleavage is slowed. Because these inhibitors bind preferentially at the extracellular surface of the transporter, their effects must result from a conformational change rather than from steric hindrance. A conformational change must also be responsible for the effect of the physiological substrate D-glucose, which is to increase the rate of cleavage. The regions of the protein involved in the conformational changes include both of the large cytoplasmic regions that are cleaved by trypsin: these are the central hydrophilic region of the sequence (residues 213-269) and the hydrophilic C-terminal region (residues 457-492).

MeSH Terms
Erythrocytes/metabolism Glucose/analogs & derivatives,metabolism Humans Monosaccharide Transport Proteins/metabolism Phloretin/pharmacology Protein Conformation Trypsin/metabolism
Chemicals
Monosaccharide Transport Proteins ethylidene glucose Trypsin Glucose Phloretin
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Gibbs A F
Department of Biochemistry and Chemistry, Royal Free Hospital School of Medicine (University of London), U.K.
Chapman D
Baldwin S A
References (41)
41 references, click to expand
  1. Kinetic properties of the reconstituted glucose transporter from human erythrocytes.
    J Biol Chem. 1981 Sep 10;256(17):8907-14 PMID: 6455434
  2. The monosaccharide transporter of the human erythrocyte. Transport activity upon reconstitution.
    J Biol Chem. 1981 Apr 25;256(8):3685-9 PMID: 7194337
  3. Changes in the intrinsic fluorescence of the human erythrocyte monosaccharide transporter upon ligand binding.
    Biochemistry. 1982 Apr 13;21(8):1905-8 PMID: 7200802
  4. Properties of N-maleoylmethionine sulphone, a novel impermeant maleimide, and its use in the selective labelling of the erythrocyte glucose-transport system.
    Biochem J. 1982 Jul 1;205(1):139-45 PMID: 7126174
  5. Monosaccharide transporter of the human erythrocyte. Characterization of an improved preparation.
    Biochemistry. 1982 Aug 3;21(16):3836-42 PMID: 6890381
  6. Endoglycosidase f cleaves the oligosaccharides from the glucose transporter of the human erythrocyte.
    Biochim Biophys Acta. 1984 Jan 25;769(2):404-10 PMID: 6421318
  7. Proteolytic and chemical dissection of the human erythrocyte glucose transporter.
    Biochem J. 1984 Jul 1;221(1):179-88 PMID: 6431970
  8. Proteolytic cleavages of cytochalasin B binding components of band 4.5 proteins of the human red blood cell membrane.
    Biochim Biophys Acta. 1984 Sep 19;776(1):10-20 PMID: 6541055
  9. Rapid kinetics of the glucose transporter from human erythrocytes. Detection and measurement of a half-turnover of the purified transporter.
    J Biol Chem. 1985 Apr 25;260(8):4575-8 PMID: 4039316
  10. Asymmetrical binding of phloretin to the glucose transport system of human erythrocytes.
    J Membr Biol. 1985;83(1-2):71-80 PMID: 4039758
  11. Sequence and structure of a human glucose transporter.
    Science. 1985 Sep 6;229(4717):941-5 PMID: 3839598
  12. The topology of the major band 4.5 protein component of the human erythrocyte membrane: characterization of reactive cysteine residues.
    Biochim Biophys Acta. 1985 Sep 25;819(1):83-92 PMID: 4041454
  13. Solubilization and separation of the human erythrocyte D-glucose transporter covalently and noncovalently photoaffinity-labeled with [3H]cytochalasin B.
    Proc Natl Acad Sci U S A. 1986 Jan;83(2):479-82 PMID: 3455783
  14. The kinetics of glucose transport in human red blood cells.
    Biochim Biophys Acta. 1986 May 28;857(2):146-54 PMID: 3707948
  15. Structural basis of human erythrocyte glucose transporter function in reconstituted vesicles.
    J Biol Chem. 1986 Jun 5;261(16):7101-4 PMID: 3711076
  16. Anomalous asymmetric kinetics of human red cell hexose transfer: role of cytosolic adenosine 5'-triphosphate.
    Biochemistry. 1986 Jun 17;25(12):3592-602 PMID: 3718945
  17. Structural basis of human erythrocyte glucose transporter function in reconstituted system. Hydrogen exchange.
    J Biol Chem. 1986 Jul 15;261(20):9155-60 PMID: 3722192
  18. ATP regulation of the human red cell sugar transporter.
    J Biol Chem. 1986 Aug 25;261(24):11028-37 PMID: 3733746
  19. Kinetics of glucose transport in human erythrocytes: zero-trans efflux and infinite-trans efflux at 0 degree C.
    Biochim Biophys Acta. 1986 Nov 17;862(2):387-98 PMID: 3778899
  20. Substrate-induced conformational change of human erythrocyte glucose transporter: inactivation by alkylating reagents.
    Biochim Biophys Acta. 1987 Jan 26;896(2):287-94 PMID: 3801473
  21. Photolabelling of the hexose transporter at external and internal sites: fragmentation patterns and evidence for a conformational change.
    Biochim Biophys Acta. 1987 Mar 12;897(3):395-405 PMID: 3545294
  22. Fourier transform infrared spectroscopic study of the structure and conformational changes of the human erythrocyte glucose transporter.
    J Biol Chem. 1987 Mar 15;262(8):3502-9 PMID: 3818652
  23. Equilibrium ligand binding to the human erythrocyte sugar transporter. Evidence for two sugar-binding sites per carrier.
    J Biol Chem. 1987 Apr 25;262(12):5464-75 PMID: 3571218
  24. Structural basis of human erythrocyte glucose transporter function in proteoliposome vesicles: circular dichroism measurements.
    Proc Natl Acad Sci U S A. 1987 Jun;84(12):4113-6 PMID: 3473495
  25. Peptide-specific antibodies as probes of the orientation of the glucose transporter in the human erythrocyte membrane.
    J Biol Chem. 1987 Jul 5;262(19):9347-52 PMID: 3597413
  26. D-glucose binding increases secondary structure of human erythrocyte monosaccharide transport protein.
    Biochem Biophys Res Commun. 1987 Jun 30;145(3):1087-91 PMID: 3606595
  27. Binding of cytochalasin B to trypsin and thermolysin fragments of the human erythrocyte hexose transporter.
    Biochim Biophys Acta. 1987 Sep 3;902(3):402-5 PMID: 3620469
  28. Investigation of the structure and function of the human erythrocyte glucose transporter by proteolytic dissection.
    Biochim Biophys Acta. 1987 Dec 11;905(2):295-310 PMID: 3689782
  29. Infinite-cis kinetics support the carrier model for erythrocyte glucose transport.
    Biochemistry. 1988 Mar 8;27(5):1441-50 PMID: 3365399
  30. The action of inhibitors on the facilitated hexose transfer system in erythrocytes.
    J Physiol. 1958 Apr 30;141(2):219-32 PMID: 13539834
  31. INHIBITION OF THE GLUCOSE PERMEABILITY OF HUMAN ERYTHROCYTES BY N-ETHYL MALEIMIDE.
    J Physiol. 1963 Oct;168:644-59 PMID: 14067949
  32. Inhibition of parallel flux and augmentation of counter flux shown by transport models not involving a mobile carrier.
    J Theor Biol. 1966 Feb;10(2):301-6 PMID: 5964395
  33. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  34. Evidence for a carrier conformational change associated with sugar transport in erythrocytes.
    Biochemistry. 1971 Mar 30;10(7):1143-8 PMID: 5553320
  35. The asymmetry of the facilitated transfer system for hexoses in human red cells and the simple kinetics of a two component model.
    J Physiol. 1973 May;231(1):143-65 PMID: 4715343
  36. Human erythrocyte sugar transport. Identification of the essential residues of the sugar carrier by specific modification.
    J Biol Chem. 1974 Mar 25;249(6):1814-22 PMID: 4817966
  37. Evidence for two asymmetric conformational states in the human erythrocyte sugar-transport system.
    Biochem J. 1975 Mar;145(3):417-29 PMID: 1156368
  38. Impermeant maleimides. Identification of an exofacial component of the human erythrocyte hexose transport mechanism.
    J Biol Chem. 1976 Nov 25;251(22):7184-90 PMID: 993210
  39. Cytochalasin B and the kinetics of inhibition of biological transport: a case of asymmetric binding to the glucose carrier.
    Biochim Biophys Acta. 1978 Jul 4;510(2):339-48 PMID: 667049
  40. The monosaccharide transport system of the human erythrocyte. Solubilization and characterization on the basis of cytochalasin B binding.
    J Biol Chem. 1978 Oct 10;253(19):6923-30 PMID: 690133
  41. Equilibria and kinetics of ligand binding to the human erythrocyte glucose transporter. Evidence for an alternating conformation model for transport.
    Biochemistry. 1981 Sep 1;20(18):5108-13 PMID: 7295669
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2560421
Published
1988-12-01
Pages
421-7
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1135426
Subset
IM
External Links
PubMed source DOI: 10.1042/bj2560421 PMC Full Text View journal details
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