A method has been developed to solubilize insulin receptors from skeletal muscles. Rat hindlimb muscles were rapidly frozen in liquid nitrogen, powdered, extracted with buffered Triton X-100, and partially purified by differential centrifugation followed by wheat germ agglutinin affinity chromatography. The solubilized receptors exhibit typical curvilinear Scatchard plots in insulin binding assays: rapid, Mn2+-dependent autophosphorylation of the β-subunit on exposure to insulin as well as insulin-stimulated kinase activity toward histone H2B. Furthermore, when intact soleus muscles were incubated in phosphate-depleted medium containing Na2H[32P]PO4, addition of insulin stimulated the in situ phosphorylation of the β-subunit of the insulin receptor. The ability to rapidly and efficiently isolate insulin receptors from skeletal muscle may permit investigation of factors that modulate insulin action in this tissue.
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July 01 1984
Phosphorylation of Insulin Receptors Solubilized from Rat Skeletal Muscle
Charles F Burant;
Charles F Burant
Departments of Medicine, Medical University of South Carolina
Charleston, South Carolina
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Mary K Treutelaar;
Mary K Treutelaar
Departments of Medicine, Medical University of South Carolina
Charleston, South Carolina
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Gary E Landreth;
Gary E Landreth
Neurology, Medical University of South Carolina
Charleston, South Carolina
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Maria G Buse
Maria G Buse
Biochemistry, Medical University of South Carolina
Charleston, South Carolina
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Address reprint requests to Dr. Maria G. Buse, Department of Medicine, Medical University of South Carolina, 171 Ashley Avenue, Charleston, South Carolina 29425.
Citation
Charles F Burant, Mary K Treutelaar, Gary E Landreth, Maria G Buse; Phosphorylation of Insulin Receptors Solubilized from Rat Skeletal Muscle. Diabetes 1 July 1984; 33 (7): 704–708. https://doi.org/10.2337/diab.33.7.704
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