Involvement of Heat Shock Protein 90 in the Degradation of Mutant Insulin Receptors by the Proteasome

• Published in: The Journal of biological chemistry Vol. 273; no. 18; pp. 11183 - 11188
• Main Authors: Imamura, Takeshi, Haruta, Tetsuro, Takata, Yasumitsu, Usui, Isao, Iwata, Minoru, Ishihara, Hajime, Ishiki, Manabu, Ishibashi, Osamu, Ueno, Eiichi, Sasaoka, Toshiyasu, Kobayashi, Masashi
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.05.1998; American Society for Biochemistry and Molecular Biology
• Subjects: Animals; Antibodies - administration & dosage; Antibodies - immunology; COS Cells; Cysteine Endopeptidases - metabolism; Endoplasmic Reticulum - metabolism; HSP90 Heat-Shock Proteins - immunology; HSP90 Heat-Shock Proteins - metabolism; Humans; Hydrolysis; Immunohistochemistry; Microinjections; Multienzyme Complexes - metabolism; Mutation; Proteasome Endopeptidase Complex; Protein Binding; Receptor, Insulin - genetics; Receptor, Insulin - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.273.18.11183

We previously reported three families with type A insulin-resistant syndrome who had mutations, either Asp1179 or Leu1193, in the kinase domain of the insulin receptor. The extreme insulin resistance of these patients was found to be caused by the decreased number of insulin receptors on the cell surface, due to the intracellular rapid degradation (Imamura, T., Takata, Y., Sasaoka, T., Takada, Y., Morioka, H., Haruta, T., Sawa, T., Iwanishi, M., Yang, G. H., Suzuki, Y., Hamada, J., and Kobayashi, M. (1994) J. Biol. Chem.269, 31019–31027). In the present study, we first examined whether these mutations caused rapid degradation of unprocessed proreceptors, using the exon 13 deleted mutant insulin receptors (ΔEx13-IR), which were accumulated in the endoplasmic reticulum as unprocessed proreceptors. The addition of Asp1179 or Leu1193 mutation to ΔEx13-IR caused accelerated degradation of the unprocessed ΔEx13-IR in the transfected COS-7 cells. Next, we tested whether these mutant receptors were degraded by the proteasome. Treatment with proteasome inhibitors Z-Leu-Leu-Nva-H (MG-115) or Z-Leu-Leu-Leu-H (MG-132) prevented the accelerated degradation of these mutant receptors, resulting in increased amounts of the mutant receptors in the COS-7 cells. Essentially the same results were obtained in the patient's transformed lymphocytes. Finally, we found that these mutant receptors bound to heat shock protein 90 (Hsp90). To determine whether Hsp90 played an important role in the accelerated receptor degradation, we examined the effect of anti-Hsp90 antibody on the mutant receptor degradation. The microinjection of anti-Hsp90 antibody into cells prevented the accelerated degradation of both Asp1179 and Leu1193 mutant insulin receptors. Taken together, these results suggest that Hsp90 is involved in dislocation of the mutant insulin receptors out of the endoplasmic reticulum into the cytosol, where the mutant receptors are degraded by the proteasome.