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 × 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 × 10−8 M). Patient 3 with acanthosis nigricans had a normal maximal autophosphorylation, but the ED50 shifted to the right (2.9 × 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.
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Original Articles|
February 01 1994
Enzyme-Linked Immunosorbent Assay Method for Human Autophosphorylated Insulin Receptor: Applicability to Insulin-Resistant States
Haruhiko Hagino;
Haruhiko Hagino
Hyogo Institute for Clinical Research
Akashi
Department of Internal Medicine, Kobe University School of Medicine
Kobe
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Kozui Shii;
Kozui Shii
Hyogo Institute for Clinical Research
Akashi
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Koichi Yokono;
Koichi Yokono
Department of Internal Medicine, Kobe University School of Medicine
Kobe
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Hiroshi Matsuba;
Hiroshi Matsuba
Hyogo Institute for Clinical Research
Akashi
Department of Internal Medicine, Kobe University School of Medicine
Kobe
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Masaki Yoshida;
Masaki Yoshida
Hyogo Institute for Clinical Research
Akashi
Department of Internal Medicine, Kobe University School of Medicine
Kobe
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Yoichi Hosomi;
Yoichi Hosomi
Hyogo Institute for Clinical Research
Akashi
Department of Internal Medicine, Kobe University School of Medicine
Kobe
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Yumi Okada;
Yumi Okada
Hyogo Institute for Clinical Research
Akashi
Department of Internal Medicine, Kobe University School of Medicine
Kobe
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Miyako Kishimoto;
Miyako Kishimoto
Department of Internal Medicine, Kobe University School of Medicine
Kobe
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Toshiki Hozumi;
Toshiki Hozumi
Division of Endocrinology and Metabolism, Department of Medicine, Hyogo Medical Center
Akashi, Japan
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Yoshihiko Ishida;
Yoshihiko Ishida
Division of Endocrinology and Metabolism, Department of Medicine, Hyogo Medical Center
Akashi, Japan
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Tsutomu Kazumi;
Tsutomu Kazumi
Division of Endocrinology and Metabolism, Department of Medicine, Hyogo Medical Center
Akashi, Japan
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Ryuichiro Nishimura;
Ryuichiro Nishimura
Hyogo Institute for Clinical Research
Akashi
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Masato Kasuga;
Masato Kasuga
Department of Internal Medicine, Kobe University School of Medicine
Kobe
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Shigeaki Baba
Shigeaki Baba
Hyogo Institute for Clinical Research
Akashi
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Address correspondence and reprint requests to Dr. Kozui Shii, Hyogo Institute for Clinical Research, 13–70 Kitaohji-cho, Akashi 673, Japan.
Diabetes 1994;43(2):274–280
Article history
Received:
February 11 1993
Revision Received:
October 07 1993
Accepted:
October 07 1993
PubMed:
8288051
Citation
Haruhiko Hagino, Kozui Shii, Koichi Yokono, Hiroshi Matsuba, Masaki Yoshida, Yoichi Hosomi, Yumi Okada, Miyako Kishimoto, Toshiki Hozumi, Yoshihiko Ishida, Tsutomu Kazumi, Ryuichiro Nishimura, Masato Kasuga, Shigeaki Baba; Enzyme-Linked Immunosorbent Assay Method for Human Autophosphorylated Insulin Receptor: Applicability to Insulin-Resistant States. Diabetes 1 February 1994; 43 (2): 274–280. https://doi.org/10.2337/diab.43.2.274
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