PMID: 3753652Feb 13, 1986Paper

Exofacial photoaffinity labelling of the human erythrocyte sugar transporter

Biochimica Et Biophysica Acta
G D HolmanP J Midgley

Abstract

The 4-azidosalicylate derivative of 1,3-bis(D-mannos-4'-yloxy)-2-[2-3H]propylamine (ASA-[2-3H]BMPA) has been tested as a photoaffinity label for the sugar transporter in human erythrocytes. When photolysed in the presence of intact erythrocytes, ASA-[2-3H]BMPA covalently binds to the exofacial surface of the transporter. This labelled protein appears as a broad band in the 4.5 region in sodium dodecyl sulphate (SDS)-polyacrylamide gel electrophoresis. The peak of radiolabel incorporation gives an apparent Mr of approx. 50 000 on 5-20% acrylamide gels. The binding is 80% inhibitable by 320 mM 4,6-O-ethylidene-D-glucose, by 320 mM D-glucose and by 50 microM cytochalasin B. Photoirradiation of a saturating concentration of ASA-BMPA in the presence of erythrocytes results in a 25-30% loss of D-galactose transport activity. From transport inactivation data and estimations of the amount of ASA-[2-3H]BMPA binding to the transporter it is calculated that there are approx. 220 000 exofacial hexose-transport binding sites per erythrocyte. The labelling of the transporter has been carried out using freshly drawn blood and 4-weeks-old transfusion blood. No change in the binding profile on SDS-polyacrylamide gel electrophoresis was observed...Continue Reading

References

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Jan 1, 1977·Methods in Enzymology·E V GromanL L Engel
Jan 10, 1985·Biochimica Et Biophysica Acta·P J MidgleyG D Holman
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Jun 20, 1980·Biochimica Et Biophysica Acta·G D Holman

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Citations

Oct 27, 1997·Journal of Pharmacological and Toxicological Methods·W M Li, J H McNeill
Dec 15, 2012·Journal of Carbohydrate Chemistry·Seok-Ho YuJennifer J Kohler
Jan 1, 1989·Annals of the New York Academy of Sciences·A CarruthersD L Melchior
Aug 1, 1992·Annals of the Rheumatic Diseases·J M CampbellR D Sturrock
Nov 10, 2005·Chemical Record : an Official Publication of the Chemical Society of Japan ... [et Al.]·Takenori TomohiroYasumaru Hatanaka
May 1, 1988·Journal of Cellular Physiology·J M May
Nov 15, 2000·Bioorganic & Medicinal Chemistry Letters·M HashimotoK Nabeta
Aug 18, 1988·Biochimica Et Biophysica Acta·J M May, B J Danzo

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