Demonstration of putative subunit(s) of a novel insulin receptor-associated 66 kDa GTP-binding protein, G(IR)

T. K. Varma, U. S. Singh, Satish Srivastava

Research output: Contribution to journalArticle

Abstract

A 66 kDa GTP-binding protein, G(ir), and insulin receptor (IR) were copurified from human placental membrane by DEAE-Sephacel and Wheat Germ Agglutinin (WGA)-Sepharose affinity chromatography. The WGA-fraction containing IR and G(ir) (IR-G(ir)-fraction) was phosphorylated (95 kDa IR-β and 66 kDa G(ir)) by IR-tyrosine kinase using [32P]ATP or photolabeled with [32P]8-azido-GTP (mainly 66 kDa), and was cross-linked with a bifunctional reagent, bis-[sulfosuccinimidyl] suberate (Bs3). The G(ir) cross-linked with putative subunit(s) to form a 110 kDa complex. Phosphorylation as well as 8-azido-GTP binding to G(ir) was not affected by cross-linking indicating that like other G-Proteins, G(ir) may also have subunits and that cross-linking of G(ir) with its putative subunits(s) does not block GTP-binding and phosphorylation sites.

Original languageEnglish (US)
Pages (from-to)517-523
Number of pages7
JournalBiochemistry and Molecular Biology International
Volume30
Issue number3
StatePublished - 1993

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Insulin Receptor
GTP-Binding Proteins
Demonstrations
Wheat Germ Agglutinins
Phosphorylation
Cross-Linking Reagents
Affinity chromatography
Agarose Chromatography
Guanosine Triphosphate
Affinity Chromatography
Sepharose
Adenosine Triphosphate
Binding Sites
Membranes
8-azidoguanosine triphosphate

ASJC Scopus subject areas

  • Biochemistry
  • Genetics
  • Molecular Biology

Cite this

Demonstration of putative subunit(s) of a novel insulin receptor-associated 66 kDa GTP-binding protein, G(IR). / Varma, T. K.; Singh, U. S.; Srivastava, Satish.

In: Biochemistry and Molecular Biology International, Vol. 30, No. 3, 1993, p. 517-523.

Research output: Contribution to journalArticle

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