Characterization of glucose transport activity reconstituted from heart and other tissues

• Published in: Biochimica et biophysica acta Vol. 1414; no. 1; pp. 217 - 230
• Main Authors: Wheeler, Thomas J., Cole, Darby, Hauck, Mary Anne
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 11.11.1998
• Subjects: Animals; Biological Transport; Diacylglycerol; Dipeptides - pharmacology; Erythrocytes - metabolism; Glucose - metabolism; Glucose transport; GLUT1; GLUT4; Glycerol - pharmacology; Humans; Liposome; Liposomes - chemistry; Membrane Proteins - chemistry; Monosaccharide Transport Proteins - antagonists & inhibitors; Monosaccharide Transport Proteins - chemistry; Monosaccharide Transport Proteins - metabolism; Myocardium - cytology; Myocardium - metabolism; Potassium Chloride - pharmacology; Rats; Reconstitution; Sodium Chloride - pharmacology; Trypsin - pharmacology; Verapamil - pharmacology; GLUT1; TPCK; HSP; Diacylglycerol; Liposome; DAG; Glucose transport; PC; PE; Reconstitution; Cbz; GLUT4; DMG
• ISSN: 0005-2736; 0006-3002; 1879-2642
• DOI: 10.1016/S0005-2736(98)00170-9

We examined several aspects of glucose transport reconstituted in liposomes, with emphasis on transporters of rat heart (mostly GLUT4) compared to those of human erythrocytes (GLUT1), and on effects of agents that modulate transport in intact cells. Several types of samples gave higher reconstituted activity using liposomes of egg lipids rather than soybean lipids. Diacylglycerol, proposed to activate transporters directly as part of the mechanism of insulin action, increased the intrinsic activity of heart transporters by only 25%, but increased the size of the reconstituted liposomes by 90%. The dipeptide Cbz-Gly-Phe-NH 2 inhibited GLUT4 with a K i of 0.2 mM, compared to 2.5 mM for GLUT1, which explains its preferential inhibition of insulin-stimulated glucose transport in adipocytes. Verapamil, which inhibits insulin- and hypoxia-stimulated glucose transport in muscle, had no effect on reconstituted transporters. Heart transporters had a higher K m for glucose uptake (13.4) than did GLUT1 (1.6 mM), in agreement with a recent study of GLUT1 and GLUT4 expressed in yeast and reconstituted in liposomes. Transporters reconstituted from heart and adipocytes were 40–70% inactivated by external trypsin, suggesting the presence of trypsin-sensitive sites on the cytoplasmic domain of GLUT4. NaCl and KCl both reduced reconstituted transport activity, but KCl had a much smaller effect on the size of the liposomes.