The glucose transport activity of human erythrocyte membranes. Reconstitution in phospholipid liposomes and fractionation by molecular sieve and ion exchange chromatography.
Human erythrocyte membranes, at a protein concentration of 1-2 g/l, were solubilized with 0.12 M cholate in the presence of 0.06 M phospholipid (egg yolk phospholipids or phosphatidylcholine). More than 40% of the protein was solubilized. Cholate was removed by molecular sieve chromatography, whereby liposomes formed. These liposomes exchanged D-glucose faster than L-glucose. The recovery of glucose transport activity in the reconstituted system was estimated to be higher than 16%. The liposomes were heterogenous in size, as shown by molecular sieve chromatography on Sepharose 4B, and small liposomes predominated. In liposomes formed with phosphatidylcholine, the distribution of glucose transport activity did not parallel the distribution of protein or phospholipid, and the activity was found mainly in the smallest liposomes. The proteins were incorporated mainly in the liposomes that eluted at the lowest ionic strength upon ion exchange chromatography. The glucose transport activity separated into three main peaks upon ion exchange chromatography of egg yolk phospholipid liposomes. The activity eluted at low ionic strength. The liposomes contained proteins mainly from the 3- and 4.5-regions (nomenclature according to Steck, T.L. (1974) J. Cell Biol. 62, 1-19). The activity peaks were highest in the first part of the chromatogram. The protein distribution did not coincide with the variation in activity over each peak. Therefore, it cannot be excluded that a minor component not seen in the electrophoretic analyses might be responsible for the glucose transport activity.
D001693:Biological Transport, Active; D001786:Blood Glucose / Q000378:metabolism*; D002793:Cholic Acids / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D006801:Humans; D008081:Liposomes; D008563:Membrane Lipids / Q000097:blood*; D008565:Membrane Proteins / Q000097:blood; D010743:Phospholipids / Q000097:blood* / Q000494:pharmacology
D001786:Blood Glucose; D002793:Cholic Acids; D008081:Liposomes; D008563:Membrane Lipids; D008565:Membrane Proteins; D010743:Phospholipids