Home LiteratureArticle Details
PMID: 2005882 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The tyrosine kinase encoded by the MET proto-oncogene is activated by autophosphorylation.

Molecular and cellular biology ·Vol. 11 ·No. 4 ·1991-04-00 ·Pages 1793-803

Naldini L, Vigna E, Ferracini R, Longati P, Gandino L, Prat M, Comoglio PM

Abstract

Protein tyrosine kinases are crucially involved in the control of cell proliferation. Therefore, the regulation of their activity in both normal and neoplastic cells has been under intense scrutiny. The product of the MET oncogene is a transmembrane receptorlike tyrosine kinase with a unique disulfide-linked heterodimeric structure. Here we show that the tyrosine kinase activity of the MET-encoded protein is powerfully activated by tyrosine autophosphorylation. The enhancement of activity was quantitated with a phosphorylation assay of exogenous substrates. It involved an increase in the Vmax of the enzyme-catalyzed phosphotransfer reaction. No change was observed in the Km (substrate). A causal relationship between tyrosine autophosphorylation and activation of the kinase activity was proved by (i) the kinetic agreement between autophosphorylation and kinase activation, (ii) the overlapping dose-response relationship for ATP, (iii) the specificity for ATP of the activation process, (iv) the phosphorylation of tyrosine residues only, in the Met protein, in the activation step, (v) the linear dependence of the activation from the input of enzyme assayed, and (vi) the reversal of the active state by phosphatase treatment. Autophosphorylation occurred predominantly on a single tryptic peptide, most likely via an intermolecular reaction. The structural features responsible for this positive modulation of kinase activity were all contained in the 45-kDa intracellular moiety of the Met protein.

MeSH Terms
Adenosine Triphosphate/metabolism Chromatography, High Pressure Liquid Enzyme Activation Kinetics Phosphorylation Protein-Tyrosine Kinases/genetics,metabolism Proto-Oncogene Mas Proto-Oncogene Proteins/genetics,metabolism Proto-Oncogene Proteins c-met Proto-Oncogenes Substrate Specificity
Chemicals
MAS1 protein, human Proto-Oncogene Mas Proto-Oncogene Proteins Adenosine Triphosphate Protein-Tyrosine Kinases Proto-Oncogene Proteins c-met
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Naldini L
Department of Biomedical Sciences and Human Oncology, University of Turin Medical School, Italy.
Vigna E
Ferracini R
Longati P
Gandino L
Prat M
Comoglio P M
References (56)
56 references, click to expand
  1. The kinetics of tyrosine phosphorylation by the purified epidermal growth factor receptor kinase of A-431 cells.
    J Biol Chem. 1983 Apr 10;258(7):4137-42 PMID: 6101263
  2. Mechanism of met oncogene activation.
    Cell. 1986 Jun 20;45(6):895-904 PMID: 2423252
  3. Autophosphorylation of the insulin receptor in vitro. Designation of phosphorylation sites and correlation with receptor kinase activation.
    J Biol Chem. 1986 Sep 15;261(26):11980-5 PMID: 3745176
  4. Phosphotyrosine antibodies identify the p210c-abl tyrosine kinase and proteins phosphorylated on tyrosine in human chronic myelogenous leukemia cells.
    Mol Cell Biol. 1986 May;6(5):1803-11 PMID: 2431286
  5. The activated human met gene encodes a protein tyrosine kinase.
    FEBS Lett. 1986 Dec 15;209(2):357-61 PMID: 3792554
  6. Insulin stimulation of the insulin receptor kinase can occur in the complete absence of beta subunit autophosphorylation.
    J Biol Chem. 1987 Feb 25;262(6):2861-8 PMID: 3546299
  7. Activation and suppression of pp60c-src transforming ability by mutation of its primary sites of tyrosine phosphorylation.
    Cell. 1987 Apr 10;49(1):65-73 PMID: 3103925
  8. Tyrosine phosphorylation regulates the biochemical and biological properties of pp60c-src.
    Cell. 1987 Apr 10;49(1):75-82 PMID: 3103926
  9. Cell transformation by pp60c-src mutated in the carboxy-terminal regulatory domain.
    Cell. 1987 Apr 10;49(1):83-91 PMID: 3103927
  10. Enzymatic activation of Fujinami sarcoma virus gag-fps transforming proteins by autophosphorylation at tyrosine.
    EMBO J. 1987 Mar;6(3):659-66 PMID: 3034604
  11. A thousand and one protein kinases.
    Cell. 1987 Sep 11;50(6):823-9 PMID: 3113737
  12. Sequence of MET protooncogene cDNA has features characteristic of the tyrosine kinase family of growth-factor receptors.
    Proc Natl Acad Sci U S A. 1987 Sep;84(18):6379-83 PMID: 2819873
  13. Characterization of the TPR-MET oncogene p65 and the MET protooncogene p140 protein-tyrosine kinases.
    Proc Natl Acad Sci U S A. 1988 Jan;85(1):21-5 PMID: 3277171
  14. Alteration of epidermal growth factor receptor activity by mutation of its primary carboxyl-terminal site of tyrosine self-phosphorylation.
    J Biol Chem. 1988 Mar 15;263(8):3610-7 PMID: 3346210
  15. Relationship of site-specific beta subunit tyrosine autophosphorylation to insulin activation of the insulin receptor (tyrosine) protein kinase activity.
    J Biol Chem. 1988 Apr 5;263(10):4593-601 PMID: 2832399
  16. 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
  17. Characterization of the mouse met proto-oncogene.
    Oncogene. 1988 Jun;2(6):593-9 PMID: 2838789
  18. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains.
    Science. 1988 Jul 1;241(4861):42-52 PMID: 3291115
  19. Structure of the met protein and variation of met protein kinase activity among human tumour cell lines.
    Br J Cancer. 1988 Jul;58(1):3-7 PMID: 3048352
  20. Growth factor receptor tyrosine kinases.
    Annu Rev Biochem. 1988;57:443-78 PMID: 3052279
  21. Biological activities of EGF-receptor mutants with individually altered autophosphorylation sites.
    EMBO J. 1988 Oct;7(10):3045-52 PMID: 3263271
  22. Kinetic parameters of the protein tyrosine kinase activity of EGF-receptor mutants with individually altered autophosphorylation sites.
    EMBO J. 1988 Oct;7(10):3053-60 PMID: 3181128
  23. Truncation of the human EGF receptor leads to differential transforming potentials in primary avian fibroblasts and erythroblasts.
    EMBO J. 1988 Oct;7(10):3061-71 PMID: 3263272
  24. p145, a protein with associated tyrosine kinase activity in a human gastric carcinoma cell line.
    Mol Cell Biol. 1988 Aug;8(8):3510-7 PMID: 3211149
  25. Evidence for autocrine activation of a tyrosine kinase in a human gastric carcinoma cell line.
    J Cell Biochem. 1988 Dec;38(4):229-36 PMID: 3241013
  26. Tyrosine kinase receptor indistinguishable from the c-met protein.
    Nature. 1989 May 11;339(6220):155-6 PMID: 2541345
  27. Signal transduction by allosteric receptor oligomerization.
    Trends Biochem Sci. 1988 Nov;13(11):443-7 PMID: 3075366
  28. Biosynthesis of the protein encoded by the c-met proto-oncogene.
    Oncogene. 1989 Nov;4(11):1383-8 PMID: 2554238
  29. Substrate phosphorylation catalyzed by the insulin receptor tyrosine kinase. Kinetic correlation to autophosphorylation of specific sites in the beta subunit.
    J Biol Chem. 1989 Dec 25;264(36):21557-72 PMID: 2557333
  30. Analysis of mutant forms of the c-src gene product containing a phenylalanine substitution for tyrosine 416.
    Oncogene Res. 1990;5(3):205-19 PMID: 1690870
  31. Signal transduction by receptors with tyrosine kinase activity.
    Cell. 1990 Apr 20;61(2):203-12 PMID: 2158859
  32. Protein kinase-c activation inhibits tyrosine phosphorylation of the c-met protein.
    Oncogene. 1990 May;5(5):721-5 PMID: 2111905
  33. Characterization of the Rous sarcoma virus transforming gene product.
    J Biol Chem. 1983 May 25;258(10):6344-51 PMID: 6304035
  34. Phosphorylation activates the insulin receptor tyrosine protein kinase.
    Proc Natl Acad Sci U S A. 1983 Jun;80(11):3237-40 PMID: 6574482
  35. Characterization and use of monoclonal antibodies for isolation of phosphotyrosyl proteins from retrovirus-transformed cells and growth factor-stimulated cells.
    Mol Cell Biol. 1983 Aug;3(8):1343-52 PMID: 6194425
  36. Detection and quantification of phosphotyrosine in proteins.
    Methods Enzymol. 1983;99:387-402 PMID: 6196603
  37. Close similarity of epidermal growth factor receptor and v-erb-B oncogene protein sequences.
    Nature. 1984 Feb 9-15;307(5951):521-7 PMID: 6320011
  38. Kinetic properties and sites of autophosphorylation of the partially purified insulin receptor from hepatoma cells.
    J Biol Chem. 1984 Jan 10;259(1):255-64 PMID: 6368536
  39. Detection of phosphotyrosine-containing proteins in the detergent-insoluble fraction of RSV-transformed fibroblasts by azobenzene phosphonate antibodies.
    EMBO J. 1984 Mar;3(3):483-9 PMID: 6201361
  40. 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
  41. Molecular cloning of a new transforming gene from a chemically transformed human cell line.
    Nature. 1984 Sep 6-11;311(5981):29-33 PMID: 6590967
  42. 42,000-molecular weight EGF receptor has protein kinase activity.
    Nature. 1984 Oct 4-10;311(5985):477-80 PMID: 6090943
  43. Immunoaffinity purification of the epidermal growth factor receptor. Stoichiometry of binding and kinetics of self-phosphorylation.
    J Biol Chem. 1984 Dec 10;259(23):14631-6 PMID: 6094567
  44. Protein-tyrosine kinases.
    Annu Rev Biochem. 1985;54:897-930 PMID: 2992362
  45. Activation of the pp60c-src kinase by middle T antigen binding or by dephosphorylation.
    EMBO J. 1985 Jun;4(6):1471-7 PMID: 2411538
  46. Autophosphorylation and protein kinase C phosphorylation of the epidermal growth factor receptor. Effect on tyrosine kinase activity and ligand binding affinity.
    J Biol Chem. 1985 Nov 25;260(27):14538-46 PMID: 2997213
  47. Self-phosphorylation enhances the protein-tyrosine kinase activity of the epidermal growth factor receptor.
    J Biol Chem. 1985 Nov 25;260(27):14642-7 PMID: 2997217
  48. The human met oncogene is related to the tyrosine kinase oncogenes.
    Nature. 1985 Nov 28-Dec 4;318(6044):385-8 PMID: 4069211
  49. A lysine in the ATP-binding site of P130gag-fps is essential for protein-tyrosine kinase activity.
    EMBO J. 1986 Jan;5(1):69-76 PMID: 3007119
  50. Insulin activation of insulin receptor tyrosine kinase in intact rat adipocytes. An in vitro system to measure histone kinase activity of insulin receptors activated in vivo.
    J Biol Chem. 1986 Apr 5;261(10):4691-7 PMID: 3007472
  51. Isolation of phosphorylated peptides and proteins on ion exchange papers.
    Anal Biochem. 1978 Jul 1;87(2):566-75 PMID: 210691
  52. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  53. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A.
    Anal Biochem. 1981 Apr;112(2):195-203 PMID: 6266278
  54. Tyrosine phosphorylation of insulin receptor beta subunit activates the receptor tyrosine kinase in intact H-35 hepatoma cells.
    J Biol Chem. 1986 Apr 5;261(10):4715-22 PMID: 3957914
  55. Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose.
    Cell. 1986 Jun 6;45(5):721-32 PMID: 3518947
  56. Immunological detection of proteins phosphorylated at tyrosine in cells stimulated by growth factors or transformed by retroviral-oncogene-coded tyrosine kinases.
    Eur J Biochem. 1986 Jul 15;158(2):383-91 PMID: 2426107
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.11.4.1793-1803.1991
Published
1991-04-00
Pages
1793-803
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC359847
Subset
IM
External Links
PubMed source DOI: 10.1128/mcb.11.4.1793-1803.1991 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2