The insulin receptor juxtamembrane region contains two independent tyrosine/β-turn internalization signals

Jonathan M. Backer, Steven E. Shoelson, Michael A. Weiss, Qing Xin Hua, R. Bentley Cheatham, Eva Haring, Deborah C. Cahill, Morris F. White

Research output: Contribution to journalArticle

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Abstract

We have investigated the role of tyrosine residues in the insulin receptor cytoplasmic juxtamembrane region (Tyr953 and Tyr960) during endocytosis. Analysis of the secondary structure of the juxtamembrane region by the Chou-Fasman algorithms predicts that both the sequences GPLY953 and NPEY960 form tyrosine-containing β-turns. Similarly, analysis of model peptides by 1-D and 2-D NMR show that these sequences form β-turns in solution, whereas replacement of the tyrosine residues with alanine destabilizes the β-turn. CHO cell lines were prepared expressing mutant receptors in which each tyrosine was mutated to phenylalanine or alanine, and an additional mutant contained alanine at both positions. These mutations had no effect on insulin binding or receptor autophosphorylation. Replacements with phenylalanine had no effect on the rate of [125I]insulin endocytosis, whereas single substitutions with alanine reduced [125I]insulin endocytosis by 40-50%. Replacement of both tyrosines with alanine reduced internalization by 70%. These data suggest that the insulin receptor contains two tyrosine/β-turns which contribute independently and additively to insulin-stimulated endocytosis.

Original languageEnglish (US)
Pages (from-to)831-839
Number of pages9
JournalJournal of Cell Biology
Volume118
Issue number4
StatePublished - Aug 1992
Externally publishedYes

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Insulin Receptor
Tyrosine
Alanine
Endocytosis
Insulin
Phenylalanine
CHO Cells
Cell Line
Peptides
Mutation

ASJC Scopus subject areas

  • Cell Biology

Cite this

Backer, J. M., Shoelson, S. E., Weiss, M. A., Hua, Q. X., Cheatham, R. B., Haring, E., ... White, M. F. (1992). The insulin receptor juxtamembrane region contains two independent tyrosine/β-turn internalization signals. Journal of Cell Biology, 118(4), 831-839.

The insulin receptor juxtamembrane region contains two independent tyrosine/β-turn internalization signals. / Backer, Jonathan M.; Shoelson, Steven E.; Weiss, Michael A.; Hua, Qing Xin; Cheatham, R. Bentley; Haring, Eva; Cahill, Deborah C.; White, Morris F.

In: Journal of Cell Biology, Vol. 118, No. 4, 08.1992, p. 831-839.

Research output: Contribution to journalArticle

Backer, JM, Shoelson, SE, Weiss, MA, Hua, QX, Cheatham, RB, Haring, E, Cahill, DC & White, MF 1992, 'The insulin receptor juxtamembrane region contains two independent tyrosine/β-turn internalization signals', Journal of Cell Biology, vol. 118, no. 4, pp. 831-839.
Backer, Jonathan M. ; Shoelson, Steven E. ; Weiss, Michael A. ; Hua, Qing Xin ; Cheatham, R. Bentley ; Haring, Eva ; Cahill, Deborah C. ; White, Morris F. / The insulin receptor juxtamembrane region contains two independent tyrosine/β-turn internalization signals. In: Journal of Cell Biology. 1992 ; Vol. 118, No. 4. pp. 831-839.
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