The Precise Sequence of FGF Receptor Autophosphorylation is Kinetically Driven and is Disrupted by Oncogenic Mutations

Overview

Journal Sci Signal

Specialties Cell Biology
Physiology
Science

Date 2009 Feb 20

PMID 19224897

Citations 82

Authors

Erin D Lew

Cristina M Furdui

Karen S Anderson

Joseph Schlessinger

Affiliations

Soon will be listed here.

Abstract

Autophosphorylation of the tyrosine kinase domain of fibroblast growth factor receptor 1 (FGFR1) is mediated by a sequential and precisely ordered three-stage autophosphorylation reaction. First-stage autophosphorylation of an activation loop tyrosine leads to 50- to 100-fold stimulation of kinase activity and is followed by second-stage phosphorylation of three additional tyrosine residues, which are binding sites for signaling molecules. Finally, third-stage phosphorylation of a second activation loop tyrosine leads to an additional 10-fold stimulation of FGFR1 catalytic activity. In this report, we show that sequential autophosphorylation of five tyrosines in the FGFR1 kinase domain is under kinetic control, mediated by both the amino acid sequence surrounding the tyrosines and their locations within the kinase structure, and, moreover, that phosphoryl transfer is the rate-limiting step. Furthermore, the strict order of autophosphorylation is disrupted by a glioblastoma-derived, oncogenic FGFR1 point mutation in the kinase domain. We propose that disrupted stepwise activation of tyrosine autophosphorylation caused by oncogenic and other activating FGFR mutations may lead to aberrant activation of and assembly of signaling molecules by the activated receptor.

Citing Articles

Ameloblastic Fibro-Odontoma with an FGFR1 Mutation: A Case Report.

Aouam O, Janssen N, de Leng W, Breimer G Head Neck Pathol. 2025; 19(1):19.

PMID: 39907837 PMC: 11799477. DOI: 10.1007/s12105-025-01758-2.

Decoding FGF/FGFR Signaling: Insights into Biological Functions and Disease Relevance.

Edirisinghe O, Ternier G, Alraawi Z, Kumar T Biomolecules. 2025; 14(12.

PMID: 39766329 PMC: 11726770. DOI: 10.3390/biom14121622.

Systems modeling of oncogenic G-protein and GPCR signaling reveals unexpected differences in downstream pathway activation.

Trogdon M, Abbott K, Arang N, Lande K, Kaur N, Tong M NPJ Syst Biol Appl. 2024; 10(1):75.

PMID: 39013872 PMC: 11252164. DOI: 10.1038/s41540-024-00400-1.

NEKL-4 regulates microtubule stability and mitochondrial health in ciliated neurons.

Power K, Nguyen K, Silva A, Singh S, Hall D, Rongo C J Cell Biol. 2024; 223(9).

PMID: 38767515 PMC: 11104396. DOI: 10.1083/jcb.202402006.

The intrinsic substrate specificity of the human tyrosine kinome.

Yaron-Barir T, Joughin B, Huntsman E, Kerelsky A, Cizin D, Cohen B Nature. 2024; 629(8014):1174-1181.

PMID: 38720073 PMC: 11136658. DOI: 10.1038/s41586-024-07407-y.

References

Murray B, Padrique E, Pinko C, McTigue M . Mechanistic effects of autophosphorylation on receptor tyrosine kinase catalysis: enzymatic characterization of Tie2 and phospho-Tie2. Biochemistry. 2001; 40(34):10243-53. DOI: 10.1021/bi010959e. View

Mohammadi M, Dikic I, Sorokin A, Burgess W, Jaye M, Schlessinger J . Identification of six novel autophosphorylation sites on fibroblast growth factor receptor 1 and elucidation of their importance in receptor activation and signal transduction. Mol Cell Biol. 1996; 16(3):977-89. PMC: 231080. DOI: 10.1128/MCB.16.3.977. View

Matte A, Tari L, Delbaere L . How do kinases transfer phosphoryl groups?. Structure. 1998; 6(4):413-9. DOI: 10.1016/s0969-2126(98)00043-4. View

Rand V, Huang J, Stockwell T, Ferriera S, Buzko O, Levy S . Sequence survey of receptor tyrosine kinases reveals mutations in glioblastomas. Proc Natl Acad Sci U S A. 2005; 102(40):14344-9. PMC: 1242336. DOI: 10.1073/pnas.0507200102. View

Favelyukis S, Till J, Hubbard S, Miller W . Structure and autoregulation of the insulin-like growth factor 1 receptor kinase. Nat Struct Biol. 2001; 8(12):1058-63. DOI: 10.1038/nsb721. View

Chen H, Ma J, Li W, Eliseenkova A, Xu C, Neubert T . A molecular brake in the kinase hinge region regulates the activity of receptor tyrosine kinases. Mol Cell. 2007; 27(5):717-30. PMC: 2094128. DOI: 10.1016/j.molcel.2007.06.028. View

Madhusudan , Trafny E, Xuong N, Adams J, Ten Eyck L, Taylor S . cAMP-dependent protein kinase: crystallographic insights into substrate recognition and phosphotransfer. Protein Sci. 1994; 3(2):176-87. PMC: 2142788. DOI: 10.1002/pro.5560030203. View

Mohammadi M, Schlessinger J, Hubbard S . Structure of the FGF receptor tyrosine kinase domain reveals a novel autoinhibitory mechanism. Cell. 1996; 86(4):577-87. DOI: 10.1016/s0092-8674(00)80131-2. View

Hubbard S, Mohammadi M, Schlessinger J . Autoregulatory mechanisms in protein-tyrosine kinases. J Biol Chem. 1998; 273(20):11987-90. DOI: 10.1074/jbc.273.20.11987. View

10.

Ablooglu A, Till J, Kim K, Parang K, Cole P, Hubbard S . Probing the catalytic mechanism of the insulin receptor kinase with a tetrafluorotyrosine-containing peptide substrate. J Biol Chem. 2000; 275(39):30394-8. DOI: 10.1074/jbc.M003524200. View

11.

Lew E, Bae J, Rohmann E, Wollnik B, Schlessinger J . Structural basis for reduced FGFR2 activity in LADD syndrome: Implications for FGFR autoinhibition and activation. Proc Natl Acad Sci U S A. 2007; 104(50):19802-7. PMC: 2148379. DOI: 10.1073/pnas.0709905104. View

12.

Wei L, Hubbard S, Hendrickson W, Ellis L . Expression, characterization, and crystallization of the catalytic core of the human insulin receptor protein-tyrosine kinase domain. J Biol Chem. 1995; 270(14):8122-30. DOI: 10.1074/jbc.270.14.8122. View

13.

Ablooglu A, Kohanski R . Activation of the insulin receptor's kinase domain changes the rate-determining step of substrate phosphorylation. Biochemistry. 2001; 40(2):504-13. DOI: 10.1021/bi002292m. View

14.

GRANT B, Adams J . Pre-steady-state kinetic analysis of cAMP-dependent protein kinase using rapid quench flow techniques. Biochemistry. 1996; 35(6):2022-9. DOI: 10.1021/bi952144+. View

15.

Jan A, Johnson E, Diamonti A, Carraway Iii K, Anderson K . Insights into the HER-2 receptor tyrosine kinase mechanism and substrate specificity using a transient kinetic analysis. Biochemistry. 2000; 39(32):9786-803. DOI: 10.1021/bi9924922. View

16.

Songyang Z, Carraway 3rd K, Eck M, Harrison S, Feldman R, Mohammadi M . Catalytic specificity of protein-tyrosine kinases is critical for selective signalling. Nature. 1995; 373(6514):536-9. DOI: 10.1038/373536a0. View

17.

Songyang Z, Cantley L . Recognition and specificity in protein tyrosine kinase-mediated signalling. Trends Biochem Sci. 1995; 20(11):470-5. DOI: 10.1016/s0968-0004(00)89103-3. View

18.

Hubbard S . Crystal structure of the activated insulin receptor tyrosine kinase in complex with peptide substrate and ATP analog. EMBO J. 1997; 16(18):5572-81. PMC: 1170189. DOI: 10.1093/emboj/16.18.5572. View

19.

Furdui C, Lew E, Schlessinger J, Anderson K . Autophosphorylation of FGFR1 kinase is mediated by a sequential and precisely ordered reaction. Mol Cell. 2006; 21(5):711-7. DOI: 10.1016/j.molcel.2006.01.022. View

20.

Sondhi D, Xu W, Songyang Z, Eck M, Cole P . Peptide and protein phosphorylation by protein tyrosine kinase Csk: insights into specificity and mechanism. Biochemistry. 1998; 37(1):165-72. DOI: 10.1021/bi9722960. View