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Transfection of AtT-20ins cells with GLUT-2 but not GLUT-1 confers glucose-stimulated insulin secretion. Relationship to glucose metabolism.

Publication ,  Journal Article
Hughes, SD; Quaade, C; Johnson, JH; Ferber, S; Newgard, CB
Published in: J Biol Chem
July 15, 1993

Glucose is thought to stimulate insulin release from islet beta-cells through generation of metabolic signals. In the current study we have introduced the genes encoding the facilitated glucose transporters known as GLUT-1 and GLUT-2 into AtT-20ins cells to assess their impact on glucose-stimulated insulin release and glucose metabolism. We find that transfection of AtT-20ins cells with GLUT-2, but not GLUT-1, confers glucose-stimulated insulin release in both static incubation and perifusion studies. Cells transfected with GLUT-1 have a Km for 3-O-methyl glucose uptake of 4 mM and a Vmax of 5-6 mmol/min/liter cell space. These values are increased compared to untransfected AtT-20ins cells (Km = 2 mM; Vmax = 0.5 mmol/min/liter cell space), but are less than observed in GLUT-2-transfected lines (Km = 16-17 mM; Vmax = 17-25 mmol/min/liter cell space). Despite these dramatic differences in glucose transport affinity and capacity, the rates of [5-3H]glucose usage are not different in the control and transfected lines over a range of glucose concentrations from 10 microM to 20 mM. We conclude that the specific effect of GLUT-2 on glucose-stimulated insulin release in AtT-20ins cells is not related to changes in the overall rate of glucose metabolism and may instead involve physical coupling of GLUT-2 with cellular proteins and/or structures involved in glucose signaling.

Duke Scholars

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

July 15, 1993

Volume

268

Issue

20

Start / End Page

15205 / 15212

Location

United States

Related Subject Headings

  • Transfection
  • RNA, Messenger
  • Monosaccharide Transport Proteins
  • Methylglucosides
  • Kinetics
  • Islets of Langerhans
  • Insulin Secretion
  • Insulin
  • Humans
  • Glucose Transporter Type 2
 

Citation

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Hughes, S. D., Quaade, C., Johnson, J. H., Ferber, S., & Newgard, C. B. (1993). Transfection of AtT-20ins cells with GLUT-2 but not GLUT-1 confers glucose-stimulated insulin secretion. Relationship to glucose metabolism. J Biol Chem, 268(20), 15205–15212.
Hughes, S. D., C. Quaade, J. H. Johnson, S. Ferber, and C. B. Newgard. “Transfection of AtT-20ins cells with GLUT-2 but not GLUT-1 confers glucose-stimulated insulin secretion. Relationship to glucose metabolism.J Biol Chem 268, no. 20 (July 15, 1993): 15205–12.
Hughes SD, Quaade C, Johnson JH, Ferber S, Newgard CB. Transfection of AtT-20ins cells with GLUT-2 but not GLUT-1 confers glucose-stimulated insulin secretion. Relationship to glucose metabolism. J Biol Chem. 1993 Jul 15;268(20):15205–12.
Hughes SD, Quaade C, Johnson JH, Ferber S, Newgard CB. Transfection of AtT-20ins cells with GLUT-2 but not GLUT-1 confers glucose-stimulated insulin secretion. Relationship to glucose metabolism. J Biol Chem. 1993 Jul 15;268(20):15205–15212.

Published In

J Biol Chem

ISSN

0021-9258

Publication Date

July 15, 1993

Volume

268

Issue

20

Start / End Page

15205 / 15212

Location

United States

Related Subject Headings

  • Transfection
  • RNA, Messenger
  • Monosaccharide Transport Proteins
  • Methylglucosides
  • Kinetics
  • Islets of Langerhans
  • Insulin Secretion
  • Insulin
  • Humans
  • Glucose Transporter Type 2