The insulin receptor functions normally in Chinese hamster ovary cells after truncation of the C terminus

M. G. Myers, J. M. Backer, K. Siddle, M. F. White

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66 Scopus citations

Abstract

We studied the structure and function of the human insulin receptor (IR) and a mutant which lacked the last 43 amino acids of the β-subunit (IR(Δct)). This deletion removed tyrosine (Tyr1322, Tyr1316) and threonine (Thr1336) phosphorylation sites. In Chinese hamster ovary (CHO) cells, insulin binding to the mutant receptor was normal, and [35S]methionine labeling indicated that both the IR and IR(Δct) were processed normally; however, the β-subunit of IR(Δct) was 5 kDa smaller than that of the IR. The time course of insulin-stimulated autophosphorylation of the partially purified IR(Δct) was normal, but the maximum autophosphorylation was reduced 20-30%. Tryptic phosphopeptide mapping confirmed the absence of the C-terminal phosphorylation sites and indicated that phosphorylation of the regulatory region (Tyr1146, Tyr1150, Tyr1151) occurred normally; kinase activity of the IR and IR(Δct) was activated normally by insulin-stimulated autophosphorylation. In the intact CHO cells, insulin-stimulated serine and threonine phosphorylation of the IR(Δct) was reduced 20%, suggesting that most Ser/Thr phosphorylation sites are located outside of the C terminus. During insulin stimulation, the wild-type and mutant insulin receptor activated the phosphatidylinositol 3-kinase. Moreover, insulin itself or human-specific anti-insulin receptor antibodies stimulated glycogen and DNA synthesis equally in both CHO/IR and CHO/IR(Δct) cells. These data suggest that the C terminus plays a minimal role in IR function and signal transmission in CHO cells.

Original languageEnglish (US)
Pages (from-to)10616-10623
Number of pages8
JournalJournal of Biological Chemistry
Volume266
Issue number16
StatePublished - 1991

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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