Regulation of insulin-stimulated glycogen synthase activity by overexpression of glutamine: fructose-6-phosphate amidotransferase in rat-1 fibroblasts.

Abstract:

:High glucose concentrations such as are seen in diabetes mellitus are known to have deleterious effects on cells, but the pathways by which glucose induces these effects are unknown. One hypothesis is that metabolism of glucose to glucosamine might be involved. For example, it has been shown that glucosamine is more potent than glucose in inducing insulin resistance in cultured adipocytes and in regulating the transcription of the growth factor transforming growth factor alpha in smooth muscle cells. The rate-limiting step in glucosamine synthesis is the conversion of fructose-6-phosphate to glucosamine-6-phosphate by the enzyme glutamine:fructose-6-phosphate amidotransferase. To test the hypothesis that this hexosamine biosynthesis pathway is involved in the induction of insulin resistance, we have overexpressed the enzyme glutamine:fructose-6-phosphate amidotransferase in Rat-1 fibroblasts and investigated its effects on insulin action in those cells. We electroporated Rat-1 fibroblasts with expression plasmids that did and did not contain the gene for glutamine:fructose-6-phosphate amidotransferase and measured glycogen synthase activity at varying insulin concentrations. Insulin stimulation was blunted in the glutamine:fructose-6-phosphate amidotransferase-transfected cells, resulting in decreased insulin sensitivity reflected by a rightward shift in the dose-response curve for activation of synthase (ED50 = 7.5 nM vs. 3.4 nM insulin, in glutamine:fructose-6-phosphate amidotransferase and control cells, respectively). Rat-1 fibroblasts incubated with 5.- mM glucosamine for 3 days exhibited a similar shift in the dose-response curve. The rightward shift in the dose-response curve is seen as early as 2 days after poration.(ABSTRACT TRUNCATED AT 250 WORDS)

journal_name

Diabetes

journal_title

Diabetes

authors

Crook ED,Daniels MC,Smith TM,McClain DA

doi

10.2337/diab.42.9.1289

subject

Has Abstract

pub_date

1993-09-01 00:00:00

pages

1289-96

issue

9

eissn

0012-1797

issn

1939-327X

journal_volume

42

pub_type

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