Enzyme-linked immunosorbent assay method for human autophosphorylated insulin receptor. Applicability to insulin-resistant states.
The insulin receptors from erythrocytes of 50 patients with non-insulin-dependent diabetes mellitus were tested for their ability to autophosphorylate. The assay was performed by a new enzyme-linked immunosorbent assay system that used monoclonal anti-insulin receptor antibodies absorbed to microtiter plates as a first antibody and polyclonal antiphosphotyrosine antibody as a labeled second antibody. By this assay, 3 patients were identified with defects in their insulin receptor kinase, although their defects appeared heterogeneous. Patient 1 had 85% less maximal autophosphorylation with a normal ED50 (1.6 x 10(-9) M insulin). Patient 2, who had polycystic ovary disease, had a 49.2% decrease in maximal autophosphorylation of insulin receptors, and the ED50 was shifted to the right (5.6 x 10(-8) M). Patient 3 with acanthosis nigricans had a normal maximal autophosphorylation, but the ED50 shifted to the right (2.9 x 10(-8) M). The mechanisms for the diversity detected in this assay is not known, but this technique has sufficient specificity and sensitivity to be used to screen for insulin-resistant patients who have a lack of kinase activity.
Hagino
H
H
Hyogo Institute for Clinical Research, Akashi, Japan.
Shii
K
K
Yokono
K
K
Matsuba
H
H
Yoshida
M
M
Hosomi
Y
Y
Okada
Y
Y
Kishimoto
M
M
Hozumi
T
T
Ishida
Y
Y
D000328:Adult; D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D003924:Diabetes Mellitus, Type 2 / Q000097:blood*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D004912:Erythrocytes / Q000378:metabolism*; D005260:Female; D015309:Glucose Clamp Technique; D006801:Humans; D007333:Insulin Resistance; D008297:Male; D008875:Middle Aged; D010766:Phosphorylation; D011972:Receptor, Insulin / Q000032:analysis* / Q000097:blood* / Q000378:metabolism; D012016:Reference Values; D014162:Transfection
D001786:Blood Glucose; D011972:Receptor, Insulin