Tyrosine-kinase defect of the insulin receptor in cultured fibroblasts from patients with lipoatropic diabetes.
Postbinding defects in insulin action were described previously in cultured fibroblasts from six patients with lipoatropic diabetes. To define the contribution of the insulin receptor tyrosine kinase in these defects, we studied autophosphorylation and kinase activity of lectin purified receptors from these six patients and six normal cell lines. The patients' insulin receptors, prepared by precipitation with polyethylene glycol, had normal insulin binding characteristics and autophosphorylation properties, but a 56% decrease in the tyrosine kinase activity toward an exogenous substrate. To identify more subtle qualitative defects in autophosphorylation, insulin receptors were sequentially immunoprecipitated and analyzed for their phosphoaminoacid content. The phosphorylated receptors precipitated with an antiphosphotyrosine antibody contained labeled phosphotyrosine, whereas those in the supernatant, when further precipitated with an antireceptor antibody, contained only phosphoserine. Under these conditions, the insulin-stimulated autophosphorylation of tyrosine was significantly decreased by 54% in the patient receptors compared to normal subjects' receptors. In addition, insulin-like growth factor-I stimulation of autophosphorylation of its receptor was reduced by 59% in the patients' cells compared to those from normal subjects. We conclude that fibroblasts from patients with lipoatropic diabetes have defects in the tyrosine kinase activity of their insulin and their insulin-like growth factor-I receptors that might give rise to the in vitro hormone resistance and be related to the in vivo hormone resistance that occurs in these patients.
Magré
J
J
Laboratory of Cell Biology INSERM U181, Saint Antoine Faculty of Medicine, Paris, France.
Grigorescu
F
F
Reynet
C
C
Caron
M
M
Capony
J P
JP
White
M F
MF
Picard
J
J
Mirouze
J
J
Capeau
J
J
D000293:Adolescent; D000328:Adult; D001345:Autoradiography; D001665:Binding Sites; D001667:Binding, Competitive; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D002846:Chromatography, Affinity; D003923:Diabetes Mellitus, Lipoatrophic / Q000378:metabolism*; D005260:Female; D006801:Humans; D007223:Infant; D007333:Insulin Resistance; D010766:Phosphorylation; D011233:Precipitin Tests; D011505:Protein-Tyrosine Kinases / Q000032:analysis* / Q000378:metabolism; D011972:Receptor, Insulin / Q000032:analysis* / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000032:analysis; D017451:Receptors, Somatomedin; D012694:Serine / Q000378:metabolism; D014443:Tyrosine / Q000276:immunology / Q000378:metabolism
D011956:Receptors, Cell Surface; D017451:Receptors, Somatomedin; D014443:Tyrosine; D012694:Serine; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin