Insulin-stimulated glucose transport in rat adipose cells. Modulation of transporter intrinsic activity by isoproterenol and adenosine

H. G. Joost, T. M. Weber, S. W. Cushman, Ian Simpson

Research output: Contribution to journalArticle

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Abstract

The mechanism of modulation of insulin-stimulated glucose transport activity in isolated rat adipose cells by lipolytic and antilipolytic agents has been examined. We have measured glucose transport activity in intact cells with 3-O-methylglucose and in plasma membranes with D-glucose, and the concentration of glucose transporters in plasma membranes using a cytochalasin B binding assay. In intact cells, isoproterenol reduced insulin-stimulated transport activity by 60%. This effect was lost after cooling and washing the cells with homogenization buffer, and neither the concentration of glucose transporters nor transport activity in the plasma membranes differed from control. However, treatment of cells with KCN prior to homogenization preserved the isoproterenol effect through the fractionation procedure. Plasma membranes from these cells contained un unchanged number of transporters (31 ± 7, mean ± S.E., versus 31 ± 4 pmol/mg of protein in controls) but transported glucose at a reduced rate (19 ± 6 versus 48 ± 9 pmol/mg of protein/s). Conversely, incubation of intact cells in the presence of adenosine stimulated plasma membrane glucose transport activity compared to that in the absence of adenosine (44 ± 6 versus 36 ± 6 pmol/mg of protein/s). Kinetic studies of isoproterenol-inhibited glucose transport in plasma membranes revealed a 60% decrease in V(max) (2900 ± 350 versus 7200 ± 1000 pmol/mg of protein/s) and a small increase in K(m) (15.1 ± 1 versus 13.0 ± 0.6 mM). These data indicate that modifications of glucose transport activity produced by lipolytic and antilipolytic agents in intact adipose cells can be fully retained in plasma membranes isolated under appropriate conditions. Furthermore, the effects of these agents occur through a modification of the glucose transporter intrinsic activity.

Original languageEnglish (US)
Pages (from-to)10033-10036
Number of pages4
JournalJournal of Biological Chemistry
Volume261
Issue number22
StatePublished - Dec 1 1986

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Cell membranes
Isoproterenol
Adenosine
Rats
Cell Membrane
Modulation
Insulin
Glucose
Facilitative Glucose Transport Proteins
Proteins
Cells
3-O-Methylglucose
Cytochalasin B
Fractionation
Plasma Cells
Washing
Assays
Buffers
Cooling
Kinetics

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

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title = "Insulin-stimulated glucose transport in rat adipose cells. Modulation of transporter intrinsic activity by isoproterenol and adenosine",
abstract = "The mechanism of modulation of insulin-stimulated glucose transport activity in isolated rat adipose cells by lipolytic and antilipolytic agents has been examined. We have measured glucose transport activity in intact cells with 3-O-methylglucose and in plasma membranes with D-glucose, and the concentration of glucose transporters in plasma membranes using a cytochalasin B binding assay. In intact cells, isoproterenol reduced insulin-stimulated transport activity by 60{\%}. This effect was lost after cooling and washing the cells with homogenization buffer, and neither the concentration of glucose transporters nor transport activity in the plasma membranes differed from control. However, treatment of cells with KCN prior to homogenization preserved the isoproterenol effect through the fractionation procedure. Plasma membranes from these cells contained un unchanged number of transporters (31 ± 7, mean ± S.E., versus 31 ± 4 pmol/mg of protein in controls) but transported glucose at a reduced rate (19 ± 6 versus 48 ± 9 pmol/mg of protein/s). Conversely, incubation of intact cells in the presence of adenosine stimulated plasma membrane glucose transport activity compared to that in the absence of adenosine (44 ± 6 versus 36 ± 6 pmol/mg of protein/s). Kinetic studies of isoproterenol-inhibited glucose transport in plasma membranes revealed a 60{\%} decrease in V(max) (2900 ± 350 versus 7200 ± 1000 pmol/mg of protein/s) and a small increase in K(m) (15.1 ± 1 versus 13.0 ± 0.6 mM). These data indicate that modifications of glucose transport activity produced by lipolytic and antilipolytic agents in intact adipose cells can be fully retained in plasma membranes isolated under appropriate conditions. Furthermore, the effects of these agents occur through a modification of the glucose transporter intrinsic activity.",
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Insulin-stimulated glucose transport in rat adipose cells. Modulation of transporter intrinsic activity by isoproterenol and adenosine. / Joost, H. G.; Weber, T. M.; Cushman, S. W.; Simpson, Ian.

In: Journal of Biological Chemistry, Vol. 261, No. 22, 01.12.1986, p. 10033-10036.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Insulin-stimulated glucose transport in rat adipose cells. Modulation of transporter intrinsic activity by isoproterenol and adenosine

AU - Joost, H. G.

AU - Weber, T. M.

AU - Cushman, S. W.

AU - Simpson, Ian

PY - 1986/12/1

Y1 - 1986/12/1

N2 - The mechanism of modulation of insulin-stimulated glucose transport activity in isolated rat adipose cells by lipolytic and antilipolytic agents has been examined. We have measured glucose transport activity in intact cells with 3-O-methylglucose and in plasma membranes with D-glucose, and the concentration of glucose transporters in plasma membranes using a cytochalasin B binding assay. In intact cells, isoproterenol reduced insulin-stimulated transport activity by 60%. This effect was lost after cooling and washing the cells with homogenization buffer, and neither the concentration of glucose transporters nor transport activity in the plasma membranes differed from control. However, treatment of cells with KCN prior to homogenization preserved the isoproterenol effect through the fractionation procedure. Plasma membranes from these cells contained un unchanged number of transporters (31 ± 7, mean ± S.E., versus 31 ± 4 pmol/mg of protein in controls) but transported glucose at a reduced rate (19 ± 6 versus 48 ± 9 pmol/mg of protein/s). Conversely, incubation of intact cells in the presence of adenosine stimulated plasma membrane glucose transport activity compared to that in the absence of adenosine (44 ± 6 versus 36 ± 6 pmol/mg of protein/s). Kinetic studies of isoproterenol-inhibited glucose transport in plasma membranes revealed a 60% decrease in V(max) (2900 ± 350 versus 7200 ± 1000 pmol/mg of protein/s) and a small increase in K(m) (15.1 ± 1 versus 13.0 ± 0.6 mM). These data indicate that modifications of glucose transport activity produced by lipolytic and antilipolytic agents in intact adipose cells can be fully retained in plasma membranes isolated under appropriate conditions. Furthermore, the effects of these agents occur through a modification of the glucose transporter intrinsic activity.

AB - The mechanism of modulation of insulin-stimulated glucose transport activity in isolated rat adipose cells by lipolytic and antilipolytic agents has been examined. We have measured glucose transport activity in intact cells with 3-O-methylglucose and in plasma membranes with D-glucose, and the concentration of glucose transporters in plasma membranes using a cytochalasin B binding assay. In intact cells, isoproterenol reduced insulin-stimulated transport activity by 60%. This effect was lost after cooling and washing the cells with homogenization buffer, and neither the concentration of glucose transporters nor transport activity in the plasma membranes differed from control. However, treatment of cells with KCN prior to homogenization preserved the isoproterenol effect through the fractionation procedure. Plasma membranes from these cells contained un unchanged number of transporters (31 ± 7, mean ± S.E., versus 31 ± 4 pmol/mg of protein in controls) but transported glucose at a reduced rate (19 ± 6 versus 48 ± 9 pmol/mg of protein/s). Conversely, incubation of intact cells in the presence of adenosine stimulated plasma membrane glucose transport activity compared to that in the absence of adenosine (44 ± 6 versus 36 ± 6 pmol/mg of protein/s). Kinetic studies of isoproterenol-inhibited glucose transport in plasma membranes revealed a 60% decrease in V(max) (2900 ± 350 versus 7200 ± 1000 pmol/mg of protein/s) and a small increase in K(m) (15.1 ± 1 versus 13.0 ± 0.6 mM). These data indicate that modifications of glucose transport activity produced by lipolytic and antilipolytic agents in intact adipose cells can be fully retained in plasma membranes isolated under appropriate conditions. Furthermore, the effects of these agents occur through a modification of the glucose transporter intrinsic activity.

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