Autophosphorylation of FGFR1 Kinase Is Mediated by a Sequential and Precisely Ordered Reaction

Furdui, Cristina M.; Lew, Erin D.; Schlessinger, Joseph; Anderson, Karen S.

doi:10.1016/j.molcel.2006.01.022

Cited by 218 publications

(223 citation statements)

References 28 publications

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“…It is noteworthy that the A-loop tyrosine in Eph RTKs is also a suboptimal phosphorylation site compared with juxtamembrane sites (13). Our data differ from the data on sequential phosphorylation of the FGFR1 kinase showing that phosphorylation on the A-loop Y653 is completed before the kinase insert and juxtamembrane sites are phosphorylated (20). This discrepancy could be because the FGFR1 kinase domain used by Furdui et al lacks the C-terminal Y766, the equivalent of Y769 in FGFR2.…”

Section: Order Of Transphosphorylation Reactionscontrasting

confidence: 92%

A crystallographic snapshot of tyrosine trans -phosphorylation in action

Chen¹,

Ma³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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Tyrosine trans-phosphorylation is a key event in receptor tyrosine kinase signaling, yet, the structural basis for this process has eluded definition. Here, we present the crystal structure of the FGF receptor 2 kinases caught in the act of trans-phosphorylation of Y769, the major C-terminal phosphorylation site. The structure reveals that enzyme-and substrate-acting kinases engage each other through elaborate and specific interactions not only in the immediate vicinity of Y769 and the enzyme active site, but also in regions that are as much of 18 Å away from D626, the catalytic base in the enzyme active site. These interactions lead to an unprecedented level of specificity and precision during the trans-phosphorylation on Y769. Time-resolved mass spectrometry analysis supports the observed mechanism of trans-phosphorylation. Our data provide a molecular framework for understanding the mechanism of action of Kallmann syndrome mutations and the order of trans-phosphorylation reactions in FGFRs. We propose that the salient mechanistic features of Y769 transphosphorylation are applicable to trans-phosphorylation of the equivalent major phosphorylation sites in many other RTKs.crystal structure ͉ FGF receptor ͉ RTKs S ignaling by receptor tyrosine kinases (RTKs) plays ubiquitous roles throughout the human life cycle commencing at germ cell maturation and continuing throughout embryogenesis into adulthood (1). Ligand binding to the extracellular region of RTKs triggers activation of the intracellular tyrosine kinase domain through the universal process of trans-phosphorylation, whereby one kinase acts as a substrate for another one. Trans-phosphorylation has two major roles in RTK signaling: trans-phosphorylation on A-loop tyrosines elevates enzyme activity while trans-phosphorylation of juxtamembrane and C-terminal tyrosines generate platforms for recruitment and phosphorylation of target substrates (2-6).Structural studies of tyrosine kinase domains have been instrumental in shaping our current understanding of the mechanisms of tyrosine kinase regulation. Crystal structures of unphosphorylated tyrosine kinase domains have unveiled diverse tactics used by kinases to achieve self-inhibition (7-10). The crystal structures of phosphorylated kinases, however, reveal how tyrosine phosphorylation stabilizes the active conformation of the kinase, and how peptide substrates dock into the enzyme active site (7,(11)(12)(13). In contrast, the structural basis for tyrosine trans-phosphorylation has remained elusive. Here, we report the crystal structure of a phosphorylated FGF receptor 2 (FGFR2) kinase domain, which provides the first crystallographic snapshot of trans-phosphorylation in action. Our structural data supported by biochemical data show that trans-phosphorylation proceeds with a much higher degree of specificity than that currently perceived based on the crystal structures of kinases in complexes with short peptide substrates. Results and DiscussionWe recently reported the crystal structure of the A-loop phosphory...

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Section: Order Of Transphosphorylation Reactionscontrasting

confidence: 92%

A crystallographic snapshot of tyrosine trans -phosphorylation in action

Chen¹,

Ma³

et al. 2008

Proc. Natl. Acad. Sci. U.S.A.

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“…Binding of FGF ligands to FGFRs induces dimerization and juxtaposition of the tyrosine kinase domains to initiate the sequential transphosphorylation of at least six tyrosine residues (Furdui et al 2006;Goetz and Mohammadi 2013;Ornitz and Itoh 2015). Activation of the FGFR tyrosine kinase domain allows the direct phosphorylation of the docked adaptor protein FGFR substrate 2α (FRS2α) and binding of other adaptor proteins, including phospholipase Cγ (PLCγ), signal transducer and activator of transcription 1 (STAT1), STAT3, and STAT5 (Ornitz and Itoh 2015).…”

Section: Fgf Signalingmentioning

confidence: 99%

Fibroblast growth factor signaling in skeletal development and disease

2015

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Fibroblast growth factor (FGF) signaling pathways are essential regulators of vertebrate skeletal development. FGF signaling regulates development of the limb bud and formation of the mesenchymal condensation and has key roles in regulating chondrogenesis, osteogenesis, and bone and mineral homeostasis. This review updates our review on FGFs in skeletal development published in Genes & Development in 2002, examines progress made on understanding the functions of the FGF signaling pathway during critical stages of skeletogenesis, and explores the mechanisms by which mutations in FGF signaling molecules cause skeletal malformations in humans. Links between FGF signaling pathways and other interacting pathways that are critical for skeletal development and could be exploited to treat genetic diseases and repair bone are also explored.

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“…Other phosphotyrosines serve as binding sites for recruitment and activation of downstream signaling molecules (Eswarakumar et al, 2005). It has recently been shown that tyrosines in FGFR1 are autophosphorylated in a sequential and strictly ordered manner (Furdui et al, 2006). In this way, the activated-FGFR transduces signals by activating different signaling cascades including Ras/mitogen-activated protein kinase (MAPK), PI3K/Akt, PLCg/PKC and p38 MAPK (Boilly et al, 2000).…”

Section: Introductionmentioning

confidence: 99%

Indirubin-3′-monoxime inhibits autophosphorylation of FGFR1 and stimulates ERK1/2 activity via p38 MAPK

et al. 2007

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Indirubin-3 0 -monoxime is a derivative of the bis-indole alkaloid indirubin, an active ingredient of a traditional Chinese medical preparation that exhibits anti-inflammatory and anti-leukemic activities. Indirubin-3 0 -monoxime is mainly recognized as an inhibitor of cyclin-dependent kinases (CDKs) and glycogen synthase kinase-3. It inhibits proliferation of cultured cells, mainly through arresting the cells in the G1/S or G2/M phase of the cell cycle. Here, we report that indirubin-3 0 -monoxime is able to inhibit proliferation of NIH/3T3 cells by specifically inhibiting autophosphorylation of fibroblast growth factor receptor 1 (FGFR1), blocking in this way the receptormediated cell signaling. Indirubin-3 0 -monoxime inhibits the activity of FGFR1 at a concentration lower than that required for inhibition of phosphorylation of CDK2 and retinoblastoma protein and cell proliferation stimulated by fetal calf serum. The ability of indirubin-3 0 -monoxime to inhibit FGFR1 signaling was similar to that of the FGFR1 inhibitor SU5402. In addition, we found that indirubin-3 0 -monoxime activates long-term p38 mitogen-activated protein kinase activity, which stimulates extracellular signal-regulated kinase 1/2 in a way unrelated to the activity of FGFR1. Furthermore, we show that indirubin-3 0 -monoxime can inhibit proliferation of the myeloid leukemia cell line KG-1a through inhibition of the activity of the FGFR1 tyrosine kinase. The data presented here demonstrate previously unknown activities of indirubin-3 0 -monoxime that may have clinical implications.

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Autophosphorylation of FGFR1 Kinase Is Mediated by a Sequential and Precisely Ordered Reaction

Cited by 218 publications

References 28 publications

A crystallographic snapshot of tyrosine trans -phosphorylation in action

A crystallographic snapshot of tyrosine trans -phosphorylation in action

Fibroblast growth factor signaling in skeletal development and disease

Indirubin-3′-monoxime inhibits autophosphorylation of FGFR1 and stimulates ERK1/2 activity via p38 MAPK

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