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/ http://www.ebi.ac.uk/ selenko-2003-2 in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Selenko P., Gregorovic G., Sprangers R., Stier G., Rhani Z., Krämer A., Sattler M. sattler@embl.de Molecular cell 2003 Only protein-protein interactions imex curation imex curation selenko-2003-1 in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Selenko P., Gregorovic G., Sprangers R., Stier G., Rhani Z., Krämer A., Sattler M. sattler@embl.de Molecular cell 2003 Only protein-protein interactions imex curation imex curation sf01_human Splicing factor 1 Zinc finger protein 162 Transcription factor ZFM1 Zinc finger gene in MEN1 locus Mammalian branch point-binding protein SF1 ZFM1 ZNF162 protein protein human Homo sapiens Human MATGANATPLDFPSKKRKRSRWNQDTMEQKTVIPGMPTVIPPGLTREQERAYIVQLQIEDLTRKLRTGDLGIPPNPEDRSPSPEPIYNSEGKRLNTREFRTRKKLEEERHNLITEMVALNPDFKPPADYKPPATRVSDKVMIPQDEYPEINFVGLLIGPRGNTLKNIEKECNAKIMIRGKGSVKEGKVGRKDGQMLPGEDEPLHALVTANTMENVKKAVEQIRNILKQGIETPEDQNDLRKMQLRELARLNGTLREDDNRILRPWQSSETRSITNTTVCTKCGGAGHIASDCKFQRPGDPQSAQDKARMDKEYLSLMAELGEAPVPASVGSTSGPATTPLASAPRPAAPANNPPPPSLMSTTQSRPPWMNSGPSESRPYHGMHGGGPGGPGGGPHSFPHPLPSLTGGHGGHPMQHNPNGPPPPWMQPPPPPMNQGPHPPGHHGPPPMDQYLGSTPVGSGVYRLHQGKGMMPPPPMGMMPPPPPPPSGQPPPPPSGPLPPWQQQQQQPPPPPPPSSSMASSTPLPWQQNTTTTTTSAGTGSIPPWQQQQAAAAASPGAPQMQGNPTMVPLPPGVQPPLPPGAPPPPPPPPPGSAGMMYAPPPPPPPPMDPSNFVTMMGMGVAGMPPFGMPPAPPPPPPQN u2af2_human Splicing factor U2AF 65 kDa subunit U2AF2 U2AF65 U2 snRNP auxiliary factor large subunit U2 auxiliary factor 65 kDa subunit protein protein human Homo sapiens Human MSDFDEFERQLNENKQERDKENRHRKRSHSRSRSRDRKRRSRSRDRRNRDQRSASRDRRRRSKPLTRGAKEEHGGLIRSPRHEKKKKVRKYWDVPPPGFEHITPMQYKAMQAAGQIPATALLPTMTPDGLAVTPTPVPVVGSQMTRQARRLYVGNIPFGITEEAMMDFFNAQMRLGGLTQAPGNPVLAVQINQDKNFAFLEFRSVDETTQAMAFDGIIFQGQSLKIRRPHDYQPLPGMSENPSVYVPGVVSTVVPDSAHKLFIGGLPNYLNDDQVKELLTSFGPLKAFNLVKDSATGLSKGYAFCEYVDINVTDQAIAGLNGMQLGDKKLLVQRASVGAKNATLVSPPSTINQTPVTLQVPGLMSSQVQMGGHPTEVLCLMNMVLPEELLDDEEYEEIVEDVRDECSKYGLVKSIEIPRPVDGVEVPGCGKIFVEFTSVFDCQKAMQGLTGRKFANRVVVTKYCDPDSYHRRDFW sf1-u2af2-17 3582963 N/A 3582965 unspecified role unspecified role prey prey purified purified N-terminal half sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false His6 tag 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 false ecobd Escherichia coli (strain B / BL21-DE3) N/A 3582966 unspecified role unspecified role bait bait purified purified RRM3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false GST tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobd Escherichia coli (strain B / BL21-DE3) N/A 3582965 competitor competitor competitor unspecified role unspecified role purified purified Binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false His6 tag 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 false chemical synthesis Chemical synthesis (Chemical synthesis) 3582975 3582972 direct interaction direct interaction false false false Fig.2A While the 1-25 residue SF1 construct is a potent competitor for the RRM3-SF1 interaction, a delta2-28 SF1 construct was unable to compete in a separate pull down experiment (see figure 2A of the paper). u2af2-sf1-4 3582963 N/A 3582966 unspecified role unspecified role bait bait purified purified P26368:p.Glu396_Glu397delinsGlyAla mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false P26368:p.Glu396_Glu397delinsAlaAla mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false P26368:p.Glu387_Glu388delinsArgArg mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false P26368:p.Glu396_Glu397delinsLysLys mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false P26368:p.Asp391_Glu394delinsAlaAlaAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P26368:p.Asp391_Glu394delinsArgArgLysLys mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false RRM3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false GST tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P26368:p.Phe454Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P26368:p.Phe454Tyr mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false P26368:p.Glu387_Glu388delinsGlyAla mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 3582965 unspecified role unspecified role prey prey purified purified N-terminal half sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15637:p.Lys15Glu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false Q15637:p.Arg17Glu mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15637:p.Arg19Glu mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15637:p.Arg21Lys mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15637:p.Arg21Asp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false Q15637:p.Trp22Phe mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15637:p.Trp22Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false Q15637:p.Asn23Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15637:p.Gln24Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15637:p.Lys15_Arg17delinsGluGluAsp mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false Q15637:p.Lys16_Lys18delinsAlaAlaAla mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15637:p.Lys16_Lys18delinsGluAspGlu mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 3582991 3582985 direct interaction direct interaction false false false 3582963 Fig.3D and E sf1-u2af2-16 3582964 N/A 3582966 unspecified role unspecified role neutral component neutral component purified purified 13C radiolabelled 13c label 13c label C13 13C C13 13C undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false RRM3 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 15N radiolabelled 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobd Escherichia coli (strain B / BL21-DE3) Stoichiometry: 1.0 N/A 3582965 unspecified role unspecified role neutral component neutral component purified purified Binding region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 13C radiolabelled 13c label 13c label C13 13C C13 13C undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 15N radiolabelled 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 3583010 3583007 direct interaction direct interaction false false false 3582964 Fig.2B-D, Fig.3A-C The quoted Kd value is an upper estimate, the authors claim that the true Kd<1uM.