Structural and biochemical evidence for an autoinhibitory role for tyrosine 984 in the juxtamembrane region of the insulin receptor.

Abstract

Tyrosine 984 in the juxtamembrane region of the insulin receptor, between the transmembrane helix and the cytoplasmic tyrosine kinase domain, is conserved among all insulin receptor-like proteins from hydra to humans. Crystallographic studies of the tyrosine kinase domain and proximal juxtamembrane region reveal that Tyr-984 interacts with several other conserved residues in the N-terminal lobe of the kinase domain, stabilizing a catalytically nonproductive position of alpha-helix C. Steady-state kinetics measurements on the soluble kinase domain demonstrate that replacement of Tyr-984 with phenylalanine results in a 4-fold increase in kcat in the unphosphorylated (basal state) enzyme. Moreover, mutation of Tyr-984 in the full-length insulin receptor results in significantly elevated receptor phosphorylation levels in cells, both in the absence of insulin and following insulin stimulation. These data demonstrate that Tyr-984 plays an important structural role in maintaining the quiescent, basal state of the insulin receptor. In addition, the structural studies suggest a possible target site for small molecule activators of the insulin receptor, with potential use in the treatment of noninsulin-dependent diabetes mellitus.

• Authors: Shiqing Li, Nicole D Covino, Evan G Stein, Jeffrey H Till, Stevan R Hubbard
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 278 Issue 28 Pg. 26007-14 (Jul 11 2003) ISSN: 0021-9258 [Print] United States
• PMID: 12707268 (Publication Type: Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, U.S. Gov't, P.H.S.)

Chemical References

• Tyrosine
• Receptor, Insulin

Topics

• Amino Acid Sequence
• Animals
• Binding Sites
• Cell Membrane (metabolism)
• Crystallography, X-Ray
• Diabetes Mellitus, Type 2 (metabolism)
• Humans
• Kinetics
• Models, Molecular
• Molecular Sequence Data
• Phosphorylation
• Protein Binding
• Protein Structure, Tertiary
• Receptor, Insulin (chemistry, metabolism, physiology)
• Transfection
• Tyrosine (chemistry)