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/ IgE-dependent activation of sphingosine kinases 1 and 2 and secretion of sphingosine 1-phosphate requires Fyn kinase and contributes to mast cell responses. IgE-dependent activation of sphingosine kinases 1 and 2 and secretion of sphingosine 1-phosphate requires Fyn kinase and contributes to mast cell responses. J. Biol. Chem. (0021-9258) 2006 imex curation Olivera A., Urtz N., Mizugishi K., Yamashita Y., Gilfillan AM., Furumoto Y., Gu H., Proia RL., Baumruker T., Rivera J. BioCreative - Critical Assessment of Information Extraction systems in Biology Presence of SPHK1 and SPHK2 confirmed by autoradiography oliveraa@mail.nih.gov 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Olivera A., Urtz N., Mizugishi K., Yamashita Y., Gilfillan AM., Furumoto Y., Gu H., Proia RL., Baumruker T., Rivera J. BioCreative - Critical Assessment of Information Extraction systems in Biology Presence of SPHK1 and SPHK2 confirmed by autoradiography oliveraa@mail.nih.gov Accepted 2006-AUG-25 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation IgE-dependent activation of sphingosine kinases 1 and 2 and secretion of sphingosine 1-phosphate requires Fyn kinase and contributes to mast cell responses. IgE-dependent activation of sphingosine kinases 1 and 2 and secretion of sphingosine 1-phosphate requires Fyn kinase and contributes to mast cell responses. J. Biol. Chem. (0021-9258) 2006 imex curation Olivera A., Urtz N., Mizugishi K., Yamashita Y., Gilfillan AM., Furumoto Y., Gu H., Proia RL., Baumruker T., Rivera J. BioCreative - Critical Assessment of Information Extraction systems in Biology Presence of SPHK1 and SPHK2 confirmed by autoradiography oliveraa@mail.nih.gov 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Olivera A., Urtz N., Mizugishi K., Yamashita Y., Gilfillan AM., Furumoto Y., Gu H., Proia RL., Baumruker T., Rivera J. BioCreative - Critical Assessment of Information Extraction systems in Biology Presence of SPHK1 and SPHK2 confirmed by autoradiography oliveraa@mail.nih.gov Accepted 2006-AUG-25 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation IgE-dependent activation of sphingosine kinases 1 and 2 and secretion of sphingosine 1-phosphate requires Fyn kinase and contributes to mast cell responses. IgE-dependent activation of sphingosine kinases 1 and 2 and secretion of sphingosine 1-phosphate requires Fyn kinase and contributes to mast cell responses. J. Biol. Chem. (0021-9258) 2006 imex curation Olivera A., Urtz N., Mizugishi K., Yamashita Y., Gilfillan AM., Furumoto Y., Gu H., Proia RL., Baumruker T., Rivera J. BioCreative - Critical Assessment of Information Extraction systems in Biology Presence of SPHK1 and SPHK2 confirmed by autoradiography oliveraa@mail.nih.gov 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Olivera A., Urtz N., Mizugishi K., Yamashita Y., Gilfillan AM., Furumoto Y., Gu H., Proia RL., Baumruker T., Rivera J. BioCreative - Critical Assessment of Information Extraction systems in Biology Presence of SPHK1 and SPHK2 confirmed by autoradiography oliveraa@mail.nih.gov Accepted 2006-AUG-25 by DAVET. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation IgE-dependent activation of sphingosine kinases 1 and 2 and secretion of sphingosine 1-phosphate requires Fyn kinase and contributes to mast cell responses. IgE-dependent activation of sphingosine kinases 1 and 2 and secretion of sphingosine 1-phosphate requires Fyn kinase and contributes to mast cell responses. J. Biol. Chem. (0021-9258) 2006 imex curation Olivera A., Urtz N., Mizugishi K., Yamashita Y., Gilfillan AM., Furumoto Y., Gu H., Proia RL., Baumruker T., Rivera J. BioCreative - Critical Assessment of Information Extraction systems in Biology Presence of SPHK1 and SPHK2 confirmed by autoradiography oliveraa@mail.nih.gov 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions mouse-bmm mouse bone marrow derived macrophages bmm bone marrow derived macrophages Bone marrow derived macrophages (BMM) were generated in vitro from precursor cells harvested from the femurs of C57BL/6 mice (7�8 weeks) by proliferation and differentiation in the presence of L-cell conditioned medium. as a source of CSF-1 anti bait coip anti bait coimmunoprecipitation anti bait coip predetermined predetermined participant predetermined Olivera A., Urtz N., Mizugishi K., Yamashita Y., Gilfillan AM., Furumoto Y., Gu H., Proia RL., Baumruker T., Rivera J. BioCreative - Critical Assessment of Information Extraction systems in Biology Presence of SPHK1 and SPHK2 confirmed by autoradiography oliveraa@mail.nih.gov Accepted 2006-AUG-25 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation sphk2_human Sphingosine kinase 2 SPHK2 SK2 protein protein human Homo sapiens Human MNGHLEAEEQQDQRPDQELTGSWGHGPRSTLVRAKAMAPPPPPLAASTPLLHGEFGSYPARGPRFALTLTSQALHIQRLRPKPEARPRGGLVPLAEVSGCCTLRSRSPSDSAAYFCIYTYPRGRRGARRRATRTFRADGAATYEENRAEAQRWATALTCLLRGLPLPGDGEITPDLLPRPPRLLLLVNPFGGRGLAWQWCKNHVLPMISEAGLSFNLIQTERQNHARELVQGLSLSEWDGIVTVSGDGLLHEVLNGLLDRPDWEEAVKMPVGILPCGSGNALAGAVNQHGGFEPALGLDLLLNCSLLLCRGGGHPLDLLSVTLASGSRCFSFLSVAWGFVSDVDIQSERFRALGSARFTLGTVLGLATLHTYRGRLSYLPATVEPASPTPAHSLPRAKSELTLTPDPAPPMAHSPLHRSVSDLPLPLPQPALASPGSPEPLPILSLNGGGPELAGDWGGAGDAPLSPDPLLSSPPGSPKAALHSPVSEGAPVIPPSSGLPLPTPDARVGASTCGPPDHLLPPLGTPLPPDWVTLEGDFVLMLAISPSHLGADLVAAPHARFDDGLVHLCWVRSGISRAALLRLFLAMERGSHFSLGCPQLGYAAARAFRLEPLTPRGVLTVDGEQVEYGPLQAQMHPGIGTLLTGPPGCPGREP F73FFCEC930DA50F sphk2_mouse Sphingosine kinase 2 Sphk2 protein protein mouse Mus musculus Mouse MAPPPLLPVAASTPILHGEFGSYPANGPRFALTLTTQALHIQRLRPKPEARPRDGLVSLDEVSGCGTLQSRSPEDTAAYFCIYTYPRGRRGGRRRATRTFRADGATTYEENRAEAQRWATALTCLLRGVPLSGDQEITPELLPRKPRLLILVNPFGGRGLAWQRCMDHVVPMISEAGLSFNLIQTERQNHARELVQGLSLSEWEGIVTVSGDGLLYEVLNGLLDRPDWEDAVRMPIGVLPCGSGNALAGAVNHHGGFEQVVGVDLLLNCSLLLCRGGSHPLDLLSVTLASGSRCFSFLSVAWGFLSDVDIHSERFRALGSARFTLGAVLGLASLHTYRGRLSYLPATTEPALPIPGHSLPRAKSELVLAPAPAPAATHSPLHRSVSDLPLPLPQPALVSPGSPEPLPDLSLNGGGPELTGDWGGAGDAPLSPDPLLPSSPNALKTAQLSPIAEGPPEMPASSGFLPPTHSAPEASTWGPVDHLLPPLGSPLPQDWVTIEGEFVLMLGILPSHLCADLMAAPHARFDDGVVHLCWVRSGISRAALLRILLAMEHGNHFSLGCPHLGYAAARAFRLEPLTPRGLLTVDGELVEYGPIQAQVHPGLATLLTGPAGQKPQA 40EE2C2C288BE26A fyn_human Tyrosine-protein kinase Fyn p59-Fyn Proto-oncogene c-Fyn Src-like kinase FYN Proto-oncogene Syn protein protein human Homo sapiens Human MGCVQCKDKEATKLTEERDGSLNQSSGYRYGTDPTPQHYPSFGVTSIPNYNNFHAAGGQGLTVFGGVNSSSHTGTLRTRGGTGVTLFVALYDYEARTEDDLSFHKGEKFQILNSSEGDWWEARSLTTGETGYIPSNYVAPVDSIQAEEWYFGKLGRKDAERQLLSFGNPRGTFLIRESETTKGAYSLSIRDWDDMKGDHVKHYKIRKLDNGGYYITTRAQFETLQQLVQHYSERAAGLCCRLVVPCHKGMPRLTDLSVKTKDVWEIPRESLQLIKRLGNGQFGEVWMGTWNGNTKVAIKTLKPGTMSPESFLEEAQIMKKLKHDKLVQLYAVVSEEPIYIVTEYMNKGSLLDFLKDGEGRALKLPNLVDMAAQVAAGMAYIERMNYIHRDLRSANILVGNGLICKIADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELVTKGRVPYPGMNNREVLEQVERGYRMPCPQDCPISLHELMIHCWKKDPEERPTFEYLQSFLEDYFTATEPQYQPGENL DIP protein P06241 original sequence version: 195 DIP protein P06241 original sequence version: 147 DIP protein P06241 original sequence version: 164 4A1E443A4B5A0977 sphk1_mouse Sphingosine kinase 1 Acetyltransferase SPHK1 Sk1 Sphk1 protein protein mouse Mus musculus Mouse MEPVECPRGLLPRPCRVLVLLNPQGGKGKALQLFQSRVQPFLEEAEITFKLILTERKNHARELVCAEELGHWDALAVMSGDGLMHEVVNGLMERPDWETAIQKPLCSLPGGSGNALAASVNHYAGYEQVTNEDLLINCTLLLCRRRLSPMNLLSLHTASGLRLYSVLSLSWGFVADVDLESEKYRRLGEIRFTVGTFFRLASLRIYQGQLAYLPVGTVASKRPASTLVQKGPVDTHLVPLEEPVPSHWTVVPEQDFVLVLVLLHTHLSSELFAAPMGRCEAGVMHLFYVRAGVSRAALLRLFLAMQKGKHMELDCPYLVHVPVVAFRLEPRSQRGVFSVDGELMVCEAVQGQVHPNYLWMVCGSRDAPSGRDSRRGPPPEEP B791FAA58FCE3D29 lyn_human Tyrosine-protein kinase Lyn JTK8 Lck/Yes-related novel protein tyrosine kinase V-yes-1 Yamaguchi sarcoma viral related oncogene homolog p53Lyn p56Lyn LYN protein protein human Homo sapiens Human MGCIKSKGKDSLSDDGVDLKTQPVRNTERTIYVRDPTSNKQQRPVPESQLLPGQRFQTKDPEEQGDIVVALYPYDGIHPDDLSFKKGEKMKVLEEHGEWWKAKSLLTKKEGFIPSNYVAKLNTLETEEWFFKDITRKDAERQLLAPGNSAGAFLIRESETLKGSFSLSVRDFDPVHGDVIKHYKIRSLDNGGYYISPRITFPCISDMIKHYQKQADGLCRRLEKACISPKPQKPWDKDAWEIPRESIKLVKRLGAGQFGEVWMGYYNNSTKVAVKTLKPGTMSVQAFLEEANLMKTLQHDKLVRLYAVVTREEPIYIITEYMAKGSLLDFLKSDEGGKVLLPKLIDFSAQIAEGMAYIERKNYIHRDLRAANVLVSESLMCKIADFGLARVIEDNEYTAREGAKFPIKWTAPEAINFGCFTIKSDVWSFGILLYEIVTYGKIPYPGRTNADVMTALSQGYRMPRVENCPDELYDIMKMCWKEKAEERPTFDYLQSVLDDFYTATEGQYQQQP DIP protein P07948 original sequence version: 141 408D3D461204E378 sphk1_human Sphingosine kinase 1 Acetyltransferase SPHK1 SK1 SPHK SPHK1 SPK protein protein human Homo sapiens Human MDPAGGPRGVLPRPCRVLVLLNPRGGKGKALQLFRSHVQPLLAEAEISFTLMLTERRNHARELVRSEELGRWDALVVMSGDGLMHEVVNGLMERPDWETAIQKPLCSLPAGSGNALAASLNHYAGYEQVTNEDLLTNCTLLLCRRLLSPMNLLSLHTASGLRLFSVLSLAWGFIADVDLESEKYRRLGEMRFTLGTFLRLAALRTYRGRLAYLPVGRVGSKTPASPVVVQQGPVDAHLVPLEEPVPSHWTVVPDEDFVLVLALLHSHLGSEMFAAPMGRCAAGVMHLFYVRAGVSRAMLLRLFLAMEKGRHMEYECPYLVYVPVVAFRLEPKDGKGVFAVDGELMVSEAVQGQVHPNYFWMVSGCVEPPPSWKPQQMPPPEEPL EB04A7F2034C2DB0 fyn_mouse Tyrosine-protein kinase Fyn p59-Fyn Fyn Proto-oncogene c-Fyn protein protein mouse Mus musculus Mouse MGCVQCKDKEAAKLTEERDGSLNQSSGYRYGTDPTPQHYPSFGVTSIPNYNNFHAAGGQGLTVFGGVNSSSHTGTLRTRGGTGVTLFVALYDYEARTEDDLSFHKGEKFQILNSSEGDWWEARSLTTGETGYIPSNYVAPVDSIQAEEWYFGKLGRKDAERQLLSFGNPRGTFLIRESETTKGAYSLSIRDWDDMKGDHVKHYKIRKLDNGGYYITTRAQFETLQQLVQHYSERAAGLCCRLVVPCHKGMPRLTDLSVKTKDVWEIPRESLQLIKRLGNGQFGEVWMGTWNGNTKVAIKTLKPGTMSPESFLEEAQIMKKLKHDKLVQLYAVVSEEPIYIVTEYMSKGSLLDFLKDGEGRALKLPNLVDMAAQVAAGMAYIERMNYIHRDLRSANILVGNGLICKIADFGLARLIEDNEYTARQGAKFPIKWTAPEAALYGRFTIKSDVWSFGILLTELVTKGRVPYPGMNNREVLEQVERGYRMPCPQDCPISLHELMIHCWKKDPEERPTFEYLQGFLEDYFTATEPQYQPGENL 99558702596DAEE0 sphk1-fyn-1 3 10 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 7 unspecified role unspecified role bait bait direct interaction direct interaction 5A We further confirmed the interaction of SphK1 with Fyn by co-immunoprecipitation analysis using purified human recombinant Fyn and [35S]methionine-labeled SphK1, transcribed/translated in an in vitro TNT T7 reticulocyte system (Fig. 5A, upper panel). sphk2-fyn-1 3 7 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction 5A Fyn interacts with SphK1 and SphK2. sphk2-lyn-1 3 9 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction 5A As shown in Fig. 5A (lower panel), an antibody to Lyn (lanes 1�3) co-immunoprecipitated [35S]methionine-labeled SphK2 only when both purified human recombinant Lyn and [35S]methionine-labeled SphK2, which was in vitro transcribed/translated, were present. sphk1-fyn-2 2 8 unspecified role unspecified role prey prey region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait direct interaction direct interaction 5B Fyn or Lyn was immunoprecipitated where indicated, and the presence of SphK1 (upper panel) or SphK2 (lower panel) was demonstrated by Western blotting using anti-V5 and anti-SphK2 (Ct), respectively. sphk1-lyn-1 2 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey region v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 5B Fyn or Lyn was immunoprecipitated where indicated, and the presence of SphK1 (upper panel) or SphK2 (lower panel) was demonstrated by Western blotting using anti-V5 and anti-SphK2 (Ct), respectively. sphk2-lyn-2 2 6 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait physical association physical association 5B Fyn or Lyn was immunoprecipitated where indicated, and the presence of SphK1 (upper panel) or SphK2 (lower panel) was demonstrated by Western blotting using anti-V5 and anti-SphK2 (Ct), respectively. sphk2-fyn-2 2 7 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 5B Fyn or Lyn was immunoprecipitated where indicated, and the presence of SphK1 (upper panel) or SphK2 (lower panel) was demonstrated by Western blotting using anti-V5 and anti-SphK2 (Ct), respectively. sphk2-fyn-3 4 11 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey direct interaction direct interaction 5C To verify that the interaction between SphK and Fyn was not an artifact of the in vitro system or the overexpression of these proteins in cells, we tested the association of endogenous SphK2 (Fig. 5C), which was the weaker interacting SphK, with Fyn in BMMC from WT and Fyn-deficient mice. FcRI sphk2-fyn-4 1 6 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. 11 unspecified role unspecified role bait bait mouse-bmm mouse bone marrow derived macrophages bmm bone marrow derived macrophages Bone marrow derived macrophages (BMM) were generated in vitro from precursor cells harvested from the femurs of C57BL/6 mice (7�8 weeks) by proliferation and differentiation in the presence of L-cell conditioned medium. as a source of CSF-1 physical association physical association 5D Fyn was immunoprecipitated from nonstimulated or Ag-stimulated (2 min) WT BMMC lysates mixed with lysates from HEK293 cells overexpressing FLAG-SphK2. FcRI (2 min)