An improved method of reconstitution of adipocyte glucose transport activity

Robinson, F W; Blevins, Teresa; Suzuki, Kazuo; Kôno, Tetsuro

doi:10.1016/0003-2697(82)90244-5

Cited by 52 publications

(13 citation statements)

References 11 publications

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“…Incubations were undertaken at room temperature (20°C). At required times, 100,ul portions (4 x 106-5 x 106 cells) of this solution were transferred to Eppendorff tubes containing 200,ul of an oil mixture and I ml of 0°C stopping solution (0.1 mM-phloretin/1 % ethanol in 0.9% NaCl/20 mmHepes, pH 7.4) (Hankin et al, 1972;Robinson et al, 1982). Transported glucose was recovered as a pellet under the oil layer, and radioactivity was counted as previously described.…”

Section: Materials and Methods Materialsmentioning

confidence: 99%

Glucose transporters in isolated chromaffin cells. Effects of insulin and secretagogues

Delicado¹,

Portugal²

1987

Biochemical Journal

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1. Isolated chromaffin cells from bovine adrenal medulla were used to study glucose transport in a homogeneous neural tissue. 2. The affinity of glucose transporters was 1.20 + 0.52 mm by the infinite-cis technique and 1.02 + 0.09 mm by the direct transport experiments. 3. The affinity for 2-deoxyglucose of these transporters was 2.3 mm. 4. The glucose transporters, quantified by [3H]cytochalasin B binding, were 419532 ±120740 receptors/cell, which corresponds to about 7.2 + 2 pmol/mg of protein, with KD = 0.1 /LM. 5. High-affinity insulin receptors with KD = 3.95 nM were present at a density of 68400 + 7500 per cell. 6. Insulin and secretagogues increased glucose transport, raising the transporter number at the plasma membrane without changes in the affinity.

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Section: Materials and Methods Materialsmentioning

confidence: 99%

Glucose transporters in isolated chromaffin cells. Effects of insulin and secretagogues

Delicado¹,

Portugal²

1987

Biochemical Journal

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“…Reconstitution of Glucose Transport Activity from Membrane Fractions-Glucose transporter protein in plasma membranes was solubilized and reconstituted into lecithin liposomes as described previously (34,35). Initial uptake rates of D-[U-…”

Section: /Ebi Data Bank With Accession Number(s) Y17801 and Y17802mentioning

confidence: 99%

GLUT8, a Novel Member of the Sugar Transport Facilitator Family with Glucose Transport Activity

Doege¹,

Schürmann²,

Bahrenberg³

et al. 2000

Journal of Biological Chemistry

230

171

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GLUT8 is a novel glucose transporter-like protein that exhibits significant sequence similarity with the members of the sugar transport facilitator family (29.4% of amino acids identical with GLUT1). Human and mouse sequence (86.2% identical amino acids) comprise 12 putative membrane-spanning helices and several conserved motifs (sugar transporter signatures), which have previously been shown to be essential for transport activity, e.g. GRK in loop 2, PETPR in loop 6, QQLSGVN in helix 7, DRAGRR in loop 8, GWGPIPW in helix 10, and PETKG in the C-terminal tail. An expressed sequence tag (STS A005N15) corresponding with the 3 -untranslated region of GLUT8 has previously been mapped to human chromosome 9. COS-7 cells transfected with GLUT8 cDNA expressed a 42-kDa protein exhibiting specific, glucose-inhibitable cytochalasin B binding (K D ‫؍‬ 56.6 ؎ 18 nM) and reconstitutable glucose transport activity (8.1 ؎ 1.4 nmol/(mg protein ؋ 10 s) versus 1.1 ؎ 0.1 in control transfections). In human tissues, a 2.4-kilobase pair transcript was predominantly found in testis, but not in testicular carcinoma. Lower amounts of the mRNA were detected in most other tissues including skeletal muscle, heart, small intestine, and brain. GLUT8 mRNA was found in testis from adult, but not from prepubertal rats; its expression in human testis was suppressed by estrogen treatment. It is concluded that GLUT8 is a sugar transport facilitator with glucose transport activity and a hormonally regulated testicular function.Hexose transport into mammalian cells is catalyzed by the members of a small family of 45-55-kDa membrane proteins, GLUT1-GLUT5 (1-4). These hexose transporters belong to the larger family of transport facilitators, which comprises yeast hexose transporters, plant hexose-proton symporters, bacterial sugar-proton symporters (5, 6), and organic anion as well as organic cation transporters (7,8). Defining characteristics in the family of hexose transporters are the presence of 12 membrane-spanning helices and a number of conserved residues and motifs (see Fig. 3). These sugar transporter signatures have been characterized by sequence comparisons as well as by mutagenesis. Substitutions, e.g. of the conserved arginine and glutamate residues on the cytoplasmic surface (9), of tryptophan residues 388 and 412 in helix 10 and 11 (10, 11), tyrosines 146 and 292/293 in helix 4 and 7 (12, 13), glutamine 161 in helix 5 (14), and glutamine 282 (15), have been shown to markedly affect transporter function. In addition, mutagenesis experiments have implicated a motif (QLS) in helix 7 in determining the sugar recognition of GLUT1-GLUT5 (16).The known glucose transporter (GLUT) 1 isoforms differ in their expression in different tissues, in their kinetic characteristics, i.e. K m values (2), and in their substrate specificity. GLUT1 mediates glucose transport into erythrocytes and through the blood-brain barrier, and appears to provide a basal supply of glucose for most cells. GLUT2 catalyzes glucose uptake into the liver (17), and is an essent...

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“…Glucose-transporter protein was solubilized and reconstituted into lecithin liposomes as described previously [16,17]. Initial uptake rates of -[U-"%C]glucose were assayed after 10 s at a substrate concentration of 5 mM.…”

Section: Reconstitution Of Glucose-transport Activity From Membrane Fmentioning

confidence: 99%

Characterization of human glucose transporter (GLUT) 11 (encoded by SLC2A11), a novel sugar-transport facilitator specifically expressed in heart and skeletal muscle

Doege

Bocianski

Scheepers

et al. 2001

Biochemical Journal

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Human GLUT11 (encoded by the solute carrier 2A11 gene, SLC2A11) is a novel sugar transporter which exhibits significant sequence similarity with the members of the GLUT family. The amino acid sequence deduced from its cDNAs predicts 12 putative membrane-spanning helices and all the motifs (sugar-transporter signatures) that have previously been shown to be essential for sugar-transport activity. The closest relative of GLUT11 is the fructose transporter GLUT5 (sharing 41.7% amino acid identity with GLUT11). The human GLUT11 gene (SLC2A11) consists of 12 exons and is located on chromosome 22q11.2. In human tissues, a 7.2kb transcript of GLUT11 was detected exclusively in heart and skeletal muscle. Transfection of COS-7 cells with GLUT11 cDNA significantly increased the glucose-transport activity reconstituted from membrane extracts as well as the specific binding of the sugar-transporter ligand cytochalasin B. In contrast to that of GLUT4, the glucose-transport activity of GLUT11 was markedly inhibited by fructose. It is concluded that GLUT11 is a novel, muscle-specific transport facilitator that is a member of the extended GLUT family of sugar/polyol-transport facilitators.

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An improved method of reconstitution of adipocyte glucose transport activity

Cited by 52 publications

References 11 publications

Glucose transporters in isolated chromaffin cells. Effects of insulin and secretagogues

Glucose transporters in isolated chromaffin cells. Effects of insulin and secretagogues

GLUT8, a Novel Member of the Sugar Transport Facilitator Family with Glucose Transport Activity

Characterization of human glucose transporter (GLUT) 11 (encoded by SLC2A11), a novel sugar-transport facilitator specifically expressed in heart and skeletal muscle

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