title: Two FGF Receptor Kinase Molecules Act in Concert to Recruit and Transphosphorylate Phospholipase Cγ.
abstract: The molecular basis by which receptor tyrosine kinases (RTKs) recruit and phosphorylate Src Homology 2 (SH2) domain-containing substrates has remained elusive. We used X-ray crystallography, NMR spectroscopy, and cell-based assays to demonstrate that recruitment and phosphorylation of Phospholipase Cγ (PLCγ), a prototypical SH2 containing substrate, by FGF receptors (FGFR) entails formation of an allosteric 2:1 FGFR-PLCγ complex. We show that the engagement of pTyr-binding pocket of the cSH2 domain of PLCγ by the phosphorylated tail of an FGFR kinase induces a conformational change at the region past the cSH2 core domain encompassing Tyr-771 and Tyr-783 to facilitate the binding/phosphorylation of these tyrosines by another FGFR kinase in trans. Our data overturn the current paradigm that recruitment and phosphorylation of substrates are carried out by the same RTK monomer in cis and disclose an obligatory role for receptor dimerization in substrate phosphorylation in addition to its canonical role in kinase activation.
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China; Department of Biochemistry & Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Zhifeng
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Department of Chemistry, New York University, New York, NY 10012, USA.
authors.forename: William M
authors.initials: WM
authors.lastname: Marsiglia
authors.affiliation: Department of Microbiology, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Upal
authors.initials: U
authors.lastname: Basu Roy
authors.affiliation: Department of Pathology and Laboratory of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
authors.forename: Nader
authors.initials: N
authors.lastname: Rahimi
authors.affiliation: Department of Biochemistry & Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Dariush
authors.initials: D
authors.lastname: Ilghari
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China; Department of Biochemistry & Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Huiyan
authors.initials: H
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China; Department of Biochemistry & Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Huaibin
authors.initials: H
authors.lastname: Chen
authors.affiliation: Department of Biochemistry & Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Weiming
authors.initials: W
authors.lastname: Gai
authors.affiliation: Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Steven
authors.initials: S
authors.lastname: Blais
authors.affiliation: Skirball Institute of Biomolecular Medicine, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Thomas A
authors.initials: TA
authors.lastname: Neubert
authors.affiliation: Department of Microbiology, New York University School of Medicine, New York, NY 10016, USA.
authors.forename: Alka
authors.initials: A
authors.lastname: Mansukhani
authors.affiliation: Department of Chemistry, New York University, New York, NY 10012, USA.
authors.forename: Nathaniel J
authors.initials: NJ
authors.lastname: Traaseth
authors.affiliation: School of Pharmacy, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China. Electronic address: xiaokunli@wzmu.edu.cn.
authors.forename: Xiaokun
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Biochemistry & Molecular Pharmacology, New York University School of Medicine, New York, NY 10016, USA. Electronic address: moosa.mohammadi@nyumc.org.
authors.forename: Moosa
authors.initials: M
authors.lastname: Mohammadi
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D018360:Crystallography, X-Ray; D066246:ErbB Receptors / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D006868:Hydrolysis; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D019906:Nuclear Magnetic Resonance, Biomolecular; D010716:Phosphatidylinositols / Q000378:metabolism; D051966:Phospholipase C gamma / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D021381:Protein Transport; D051496:Receptor, Fibroblast Growth Factor, Type 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051497:Receptor, Fibroblast Growth Factor, Type 2 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017479:Receptors, Platelet-Derived Growth Factor / Q000378:metabolism; D013329:Structure-Activity Relationship; D014162:Transfection; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism; D018909:src Homology Domains
chemical_list: D009097:Multienzyme Complexes; D010716:Phosphatidylinositols; D066246:ErbB Receptors; C496348:FGFR1 protein, human; C496353:FGFR2 protein, human; D051496:Receptor, Fibroblast Growth Factor, Type 1; D051497:Receptor, Fibroblast Growth Factor, Type 2; D017479:Receptors, Platelet-Derived Growth Factor; D040301:Vascular Endothelial Growth Factor Receptor-2; D051966:Phospholipase C gamma; C117399:Plcg1 protein, mouse; C000607812:phospholipase Cgamma1, rat