Defect in tyrosine kinase activity of the insulin receptor from a patient with insulin resistance and acanthosis nigricans.
We report here a defect in tyrosine kinase activity of the insulin receptor from an insulin-resistant patient with acanthosis nigricans using cultured Ebstein-Barr virus (EBV)-transformed B-lymphocytes. As judged by affinity labeling and immunoblotting, the alpha- and beta-subunits of insulin receptors from the patient's lymphocytes exhibited the same mol wt as those from control subjects. Lectin-purified extracts from lymphocytes of the patient and the control subjects containing the same insulin-binding capacity were assayed for autophosphorylation and the ability to phosphorylate histone H2B. The degree of insulin-dependent autophosphorylation and the tyrosine kinase activity of the insulin receptor from the patient's lymphocytes were decreased to 15% and 13%, respectively, in a cell-free system. The insulin-dependent autophosphorylation of the insulin receptor was also impaired in intact EBV lymphocytes from the patient. Consistent with these results, we found that one of this patient's alleles had a mutation in which valine is substituted for Gly996, the third glycine in the conserved Gly-X-Gly-X-X-Gly motif in the kinase domain. Thus, it seems likely that the defect in tyrosine kinase activity of the insulin receptor cause the insulin resistance in this patient. The EBV lymphocyte can be a good system to detect genetically determined abnormalities in the insulin receptor.
Yamamoto
R
R
Third Department of Internal Medicine, Faculty of Medicine, University of Tokyo, Japan.
Shiba
T
T
Tobe
K
K
Shibasaki
Y
Y
Koshio
O
O
Izumi
T
T
Odawara
M
M
Mikami
Y
Y
Matsuura
N
N
Akanuma
Y
Y
D000052:Acanthosis Nigricans / Q000201:enzymology* / Q000235:genetics; D000255:Adenosine Triphosphate / Q000032:analysis; D000293:Adolescent; D000483:Alleles; D001402:B-Lymphocytes / Q000378:metabolism; D001665:Binding Sites; D002472:Cell Transformation, Viral; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis; D005998:Glycine; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D007333:Insulin Resistance; D008297:Male; D009154:Mutation; D010449:Peptide Mapping; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000235:genetics* / Q000378:metabolism; D011972:Receptor, Insulin / Q000235:genetics* / Q000378:metabolism; D014633:Valine
D007328:Insulin; D000255:Adenosine Triphosphate; D004247:DNA; D011505:Protein-Tyrosine Kinases; D011972:Receptor, Insulin; D014633:Valine; D005998:Glycine