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PMID: 1312712 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

YMXM motifs of IRS-1 define substrate specificity of the insulin receptor kinase.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 89 ·No. 6 ·1992-03-15 ·Pages 2027-31

Shoelson SE, Chatterjee S, Chaudhuri M, White MF

Abstract

Of 34 tyrosine residues in insulin receptor substrate 1 (IRS-1), 14 are adjacent to acidic residues, suggesting that they might be phosphorylation sites. Synthetic peptides corresponding to sequences surrounding these tyrosines were used as substrates of the insulin receptor kinase. Surprisingly six of these, each within YMXM motifs, were phosphorylated with greatest efficiency (Km, 24-92 microM; kcat/Km, 0.6-2.1 x 10(4) M-1.sec-1). Substituted YMXM peptides revealed a strong preference of the insulin receptor kinase for methionine at Y + 1 and Y + 3 positions. When phosphorylated, related YMXM sequences are recognition motifs for binding to proteins with src-homology (SH2) domains. The combined hydrophobic and flexible nature of methionine side chains adjacent to the targeted tyrosines provides a versatile contact for recognition by diverse proteins involved in signal transduction.

MeSH Terms
Amino Acid Sequence Animals CHO Cells Cricetinae Humans Kinetics Methionine Molecular Sequence Data Peptides/chemical synthesis Phosphates/metabolism Phosphorylation Protein-Tyrosine Kinases/genetics,metabolism Receptor, Insulin/genetics,metabolism Substrate Specificity Transfection
Chemicals
Peptides Phosphates Methionine Protein-Tyrosine Kinases Receptor, Insulin
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Shoelson S E
Joslin Diabetes Center, Department of Medicine, Brigham and Women's Hospital, Boston, MA.
Chatterjee S
Chaudhuri M
White M F
References (39)
39 references, click to expand
  1. Statistical analysis of enzyme kinetic data.
    Methods Enzymol. 1979;63:103-38 PMID: 502857
  2. cDNA cloning of a novel 85 kd protein that has SH2 domains and regulates binding of PI3-kinase to the PDGF beta-receptor.
    Cell. 1991 Apr 5;65(1):75-82 PMID: 1849460
  3. Phosphatidylinositol kinase or an associated protein is a substrate for the insulin receptor tyrosine kinase.
    J Biol Chem. 1990 Jan 5;265(1):396-400 PMID: 1688432
  4. SH2 and SH3 domains: elements that control interactions of cytoplasmic signaling proteins.
    Science. 1991 May 3;252(5006):668-74 PMID: 1708916
  5. Oncogenes and signal transduction.
    Cell. 1991 Jan 25;64(2):281-302 PMID: 1846320
  6. On the role of methionine residues in the sequence-independent recognition of nonpolar protein surfaces.
    Biochemistry. 1991 Jul 9;30(27):6633-6 PMID: 2065050
  7. Activation of phosphatidylinositol 3-kinase by insulin.
    Proc Natl Acad Sci U S A. 1990 Feb;87(4):1411-5 PMID: 2154747
  8. Src homology region 2 domains direct protein-protein interactions in signal transduction.
    Proc Natl Acad Sci U S A. 1990 Nov;87(21):8622-6 PMID: 2236073
  9. Insulin rapidly stimulates tyrosine phosphorylation of a Mr-185,000 protein in intact cells.
    Nature. 1985 Nov 14-20;318(6042):183-6 PMID: 2414672
  10. A cascade of tyrosine autophosphorylation in the beta-subunit activates the phosphotransferase of the insulin receptor.
    J Biol Chem. 1988 Feb 25;263(6):2969-80 PMID: 2449432
  11. Nonphosphorylatable substrate analogs selectively block autophosphorylation and activation of the insulin receptor, epidermal growth factor receptor, and pp60v-src kinases.
    J Biol Chem. 1989 May 15;264(14):7831-6 PMID: 2498306
  12. Binding kinetics of mutated insulin receptors in transfected cells grown in suspension culture: application to the Tyr----Phe 960 insulin receptor mutant.
    Biochem Biophys Res Commun. 1989 Oct 16;164(1):191-8 PMID: 2508634
  13. The human insulin receptor cDNA: the structural basis for hormone-activated transmembrane signalling.
    Cell. 1985 Apr;40(4):747-58 PMID: 2859121
  14. Human insulin receptor and its relationship to the tyrosine kinase family of oncogenes.
    Nature. 1985 Feb 28-Mar 6;313(6005):756-61 PMID: 2983222
  15. Computer analysis of enzyme-substrate-inhibitor kinetic data with automatic model selection using IBM-PC compatible microcomputers.
    Enzyme. 1986;36(3):197-206 PMID: 3803348
  16. Insulin stimulates tyrosine phosphorylation of the insulin receptor in a cell-free system.
    Nature. 1982 Aug 12;298(5875):667-9 PMID: 6178977
  17. Ligand: a versatile computerized approach for characterization of ligand-binding systems.
    Anal Biochem. 1980 Sep 1;107(1):220-39 PMID: 6254391
  18. Synthetic peptide substrates for a tyrosine protein kinase.
    J Biol Chem. 1982 May 10;257(9):4843-8 PMID: 6279650
  19. Phosphorylation of exogenous substrates by the insulin receptor-associated protein kinase.
    J Biol Chem. 1983 Jun 10;258(11):6682-5 PMID: 6406486
  20. Phosphorylation activates the insulin receptor tyrosine protein kinase.
    Proc Natl Acad Sci U S A. 1983 Jun;80(11):3237-40 PMID: 6574482
  21. Phosphorylation of gastrin-17 by epidermal growth factor-stimulated tyrosine kinase.
    Nature. 1983 Feb 3;301(5899):435-7 PMID: 6600511
  22. Phosphorylation of a synthetic gastrin peptide by the tyrosine kinase of A431 cell membranes.
    Biochem Biophys Res Commun. 1982 Dec 15;109(3):656-63 PMID: 6891588
  23. Purification and partial sequence analysis of pp185, the major cellular substrate of the insulin receptor tyrosine kinase.
    J Biol Chem. 1991 May 5;266(13):8302-11 PMID: 2022647
  24. How calmodulin binds its targets: sequence independent recognition of amphiphilic alpha-helices.
    Trends Biochem Sci. 1990 Feb;15(2):59-64 PMID: 2186516
  25. Protein kinase recognition sequence motifs.
    Trends Biochem Sci. 1990 Sep;15(9):342-6 PMID: 2238044
  26. Purification of recombinant pp60v-src protein tyrosine kinase and phosphorylation of peptides with different secondary structure preference.
    Biochemistry. 1989 Nov 14;28(23):9047-52 PMID: 2481500
  27. Model for signal sequence recognition from amino-acid sequence of 54K subunit of signal recognition particle.
    Nature. 1989 Aug 10;340(6233):482-6 PMID: 2502718
  28. Tryptic activation of the insulin receptor. Proteolytic truncation of the alpha-subunit releases the beta-subunit from inhibitory control.
    J Biol Chem. 1988 Apr 5;263(10):4852-60 PMID: 2832409
  29. Phosphorylation of synthetic insulin receptor peptides by the insulin receptor kinase and evidence that the preferred sequence containing Tyr-1150 is phosphorylated in vivo.
    J Biol Chem. 1986 Jul 25;261(21):10000-5 PMID: 3733682
  30. Expression of a functional human insulin receptor from a cloned cDNA in Chinese hamster ovary cells.
    Proc Natl Acad Sci U S A. 1985 Dec;82(23):8014-8 PMID: 3906655
  31. Polyoma middle-sized T antigen can be phosphorylated on tyrosine at multiple sites in vitro.
    EMBO J. 1984 Jan;3(1):73-9 PMID: 6200324
  32. Analysis of the sequence of amino acids surrounding sites of tyrosine phosphorylation.
    Proc Natl Acad Sci U S A. 1982 Feb;79(4):973-7 PMID: 6280176
  33. Tyrosine phosphorylation of the insulin receptor beta subunit activates the receptor-associated tyrosine kinase activity.
    J Biol Chem. 1984 Apr 25;259(8):5277-86 PMID: 6538876
  34. In vitro phosphorylation of angiotensin analogs by tyrosyl protein kinases.
    J Biol Chem. 1983 Jan 25;258(2):1022-5 PMID: 6822491
  35. Insulin stimulates the phosphorylation of the 95,000-dalton subunit of its own receptor.
    Science. 1982 Jan 8;215(4529):185-7 PMID: 7031900
  36. Isolation of phosphorylated peptides and proteins on ion exchange papers.
    Anal Biochem. 1978 Jul 1;87(2):566-75 PMID: 210691
  37. Consensus sequences as substrate specificity determinants for protein kinases and protein phosphatases.
    J Biol Chem. 1991 Aug 25;266(24):15555-8 PMID: 1651913
  38. A phosphatidylinositol-3 kinase binds to platelet-derived growth factor receptors through a specific receptor sequence containing phosphotyrosine.
    Mol Cell Biol. 1991 Feb;11(2):1125-32 PMID: 1703628
  39. Structure of the insulin receptor substrate IRS-1 defines a unique signal transduction protein.
    Nature. 1991 Jul 4;352(6330):73-7 PMID: 1648180
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.89.6.2027
Published
1992-03-15
Pages
2027-31
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC48589
Subset
IM
Grants
NIDDK NIH HHS · DK08366 · United States
External Links
PubMed source DOI: 10.1073/pnas.89.6.2027 PMC Full Text View journal details
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