IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Cell (0092-8674) 2012 imex curation Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org Accepted 2012-AUG-01 by MGT 19-Sept-2012: Contacted by IntAct-Help. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Cell (0092-8674) 2012 imex curation Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro mst microscale thermophoresis mst tag fluorescence tag visualisation by fluorescence tag fluorescence Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org Accepted 2012-AUG-01 by MGT 19-Sept-2012: Contacted by IntAct-Help. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Cell (0092-8674) 2012 imex curation Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org Accepted 2012-AUG-01 by MGT 19-Sept-2012: Contacted by IntAct-Help. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Cell (0092-8674) 2012 imex curation Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro light scattering light scattering predetermined predetermined participant predetermined Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org Accepted 2012-AUG-01 by MGT 19-Sept-2012: Contacted by IntAct-Help. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Cell (0092-8674) 2012 imex curation Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay western blot western blot Immuno blot Immuno blot Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org Accepted 2012-AUG-01 by MGT 19-Sept-2012: Contacted by IntAct-Help. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Cell (0092-8674) 2012 imex curation Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org Accepted 2012-AUG-01 by MGT 19-Sept-2012: Contacted by IntAct-Help. Cell (0092-8674) 2012 Only protein-protein interactions imex curation Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Inhibition of Basal FGF Receptor Signaling by Dimeric Grb2. Cell (0092-8674) 2012 imex curation Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org 19-Sept-2012: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Lin C.C., Melo F.A., Ghosh R., Suen K.M., Stagg L.J., Kirkpatrick J., Arold S.T., Ahmed Z., Ladbury J.E. Cancer - Interactions investigated in the context of cancer Source of FGFR2 and GRB2 obtained from the author. jeladbury@mdanderson.org Accepted 2012-AUG-01 by MGT 19-Sept-2012: Contacted by IntAct-Help. Cell (0092-8674) 2012 Only protein-protein interactions imex curation fgfr2_human Fibroblast growth factor receptor 2 FGFR2 BEK KSAM Keratinocyte growth factor receptor KGFR K-sam protein protein human Homo sapiens Human MVSWGRFICLVVVTMATLSLARPSFSLVEDTTLEPEEPPTKYQISQPEVYVAAPGESLEVRCLLKDAAVISWTKDGVHLGPNNRTVLIGEYLQIKGATPRDSGLYACTASRTVDSETWYFMVNVTDAISSGDDEDDTDGAEDFVSENSNNKRAPYWTNTEKMEKRLHAVPAANTVKFRCPAGGNPMPTMRWLKNGKEFKQEHRIGGYKVRNQHWSLIMESVVPSDKGNYTCVVENEYGSINHTYHLDVVERSPHRPILQAGLPANASTVVGGDVEFVCKVYSDAQPHIQWIKHVEKNGSKYGPDGLPYLKVLKAAGVNTTDKEIEVLYIRNVTFEDAGEYTCLAGNSIGISFHSAWLTVLPAPGREKEITASPDYLEIAIYCIGVFLIACMVVTVILCRMKNTTKKPDFSSQPAVHKLTKRIPLRRQVTVSAESSSSMNSNTPLVRITTRLSSTADTPMLAGVSEYELPEDPKWEFPRDKLTLGKPLGEGCFGQVVMAEAVGIDKDKPKEAVTVAVKMLKDDATEKDLSDLVSEMEMMKMIGKHKNIINLLGACTQDGPLYVIVEYASKGNLREYLRARRPPGMEYSYDINRVPEEQMTFKDLVSCTYQLARGMEYLASQKCIHRDLAARNVLVTENNVMKIADFGLARDINNIDYYKKTTNGRLPVKWMAPEALFDRVYTHQSDVWSFGVLMWEIFTLGGSPYPGIPVEELFKLLKEGHRMDKPANCTNELYMMMRDCWHAVPSQRPTFKQLVEDLDRILTLTTNEEYLDLSQPLEQYSPSYPDTRSSCSSGDDSVFSPDPMPYEPCLPQYPHINGSVKT 6CD5001C960ED82F grb2_human Growth factor receptor-bound protein 2 GRB2 ASH Adapter protein GRB2 SH2/SH3 adapter GRB2 Protein Ash protein protein human Homo sapiens Human MEAIAKYDFKATADDELSFKRGDILKVLNEECDQNWYKAELNGKDGFIPKNYIEMKPHPWFFGKIPRAKAEEMLSKQRHDGAFLIRESESAPGDFSLSVKFGNDVQHFKVLRDGAGKYFLWVVKFNSLNELVDYHRSTSVSRNQQIFLRDIEQVPQQPTYVQALFDFDPQEDGELGFRRGDFIHVMDNSDPNWWKGACHGQTGMFPRNYVTPVNRNV 83A4B0BA1B248DC4 fgfr2-2 5 8 putative self putative self putative self putative self kinase domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation true Supp. Fig. 1C, 1D fgfr2-grb2 5 8 putative self putative self putative self putative self c-terminal region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 stimulator stimulator unspecified role unspecified role region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Supp. Fig. 1C, 1D fgfr2-grb2-6 1 8 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined chemical synthesis Chemical synthesis (Chemical synthesis) 9 unspecified role unspecified role neutral component neutral component region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 1h region isotope label isotope label undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region 13c label 13c label C13 13C C13 13C undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined sh3 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 30 23 direct interaction direct interaction Supp. Fig. 2A grb2-1 2 9 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 2C fgfr2-grb2-3 4 9 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 2B, S3. fgfr2-grb2-4 7 8 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role neutral component neutral component region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 3A, 3B fgfr2-fgfr2-1 6 8 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role fluorescence donor fluorescence donor bfp tag region fluorescent prot tag fluorescent protein tag fluorescent prot tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 53 50 direct interaction direct interaction Fig. 3C, 4A, Supp. Fig. 3D Interaction only observed when the GRB2 dimer was present, suggesting that GRB2-binding may bring the 2 molecules together. Grb2 therefore exists as an isolated dimer in solution, which forms a 2:2 heterotetrameric complex binds with FGFR2 under saturating concentrations of receptor. grb2-grb2 3 9 unspecified role unspecified role fluorescence donor fluorescence donor region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) proximity proximity close-contact colocalization neighbourhood interaction Supp. Fig. 4 Grb2 dimers were observed to be cytoplasmically dispersed throughout cells without FGFR2, but membrane localized in the presence of FGFR2 fgfr2-grb2-5 5 8 enzyme enzyme unspecified role unspecified role cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined c_terminal_region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 enzyme target enzyme target substrate substrate unspecified role unspecified role region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined sh3 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Fig. 5A, 5B