Calmodulin antagonists inhibit insulin-stimulated GLUT4 (Glucose Transporter 4) translocation by preventing the formation of phosphatidylinositol 3,4,5-trisphosphate in 3T3L1 adipocytes

Chunmei Yang, Robert T. Watson, Jeffrey S. Elmendorf, David B. Sacks, Jeffrey E. Pessin

Research output: Contribution to journalArticle

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Abstract

It has been previously reported that calmodulin plays a regulatory role in the insulin stimulation of glucose transport. To examine the basis for this observation, we examined the effect of a panel of calmodulin antagonists that demonstrated a specific inhibition of insulin-stimulated glucose transporter 4 (GLUT4) but not insulin- or platelet-derived growth factor (PDGF)-stimulated GLUT1 translocation in 3T3L1 adipocytes. These treatments had no effect on insulin receptor autophosphorylation or tyrosine phosphorylation of insulin receptor substrate 1 (IRS1). Furthermore, IRS1 or phosphotyrosine antibody immunoprecipitation of phosphatidylinositol (PI) 3-kinase activity was not affected. Despite the marked insulin and PDGF stimulation of PI 3-kinase activity, there was a near complete inhibition of protein kinase B activation. Using a fusion protein of the Grp1 pleckstrin homology (PH) domain with the enhanced green fluorescent protein, we found that the calmodulin antagonists prevented the insulin stimulation of phosphatidylinositol 3,4,5-trisphosphate [PI(3,4,5)P3] formation in vivo. Similarly, although PDGF stimulation increased PI 3-kinase activity in in vitro immunoprecipitation assays, there was also no significant formation of PI(3,4,5)P3 in vivo. These data demonstrate that calmodulin antagonists prevent insulin-stimulated GLUT4 translocation by inhibiting the in vivo production of PI(3,4,5)P3 without directly affecting IRS1- or phosphotyrosine-associated PI 3-kinase activity. This phenomenon is similar to that observed for the PDGF stimulation of 3T3L1 adipocytes.

Original languageEnglish (US)
Pages (from-to)317-326
Number of pages10
JournalMolecular Endocrinology
Volume14
Issue number2
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

Insulin Antagonists
Phosphatidylinositol 3-Kinase
Facilitative Glucose Transport Proteins
Platelet-Derived Growth Factor
Calmodulin
Adipocytes
Insulin Receptor Substrate Proteins
Phosphatidylinositols
Insulin
Phosphotyrosine
Immunoprecipitation
Proto-Oncogene Proteins c-akt
Insulin Receptor
Tyrosine
Phosphorylation
Glucose
phosphatidylinositol 3,4,5-triphosphate
Antibodies
Proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology, Diabetes and Metabolism

Cite this

Calmodulin antagonists inhibit insulin-stimulated GLUT4 (Glucose Transporter 4) translocation by preventing the formation of phosphatidylinositol 3,4,5-trisphosphate in 3T3L1 adipocytes. / Yang, Chunmei; Watson, Robert T.; Elmendorf, Jeffrey S.; Sacks, David B.; Pessin, Jeffrey E.

In: Molecular Endocrinology, Vol. 14, No. 2, 2000, p. 317-326.

Research output: Contribution to journalArticle

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