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/ Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. Cell. Signal. (0898-6568) 2011 imex curation Ger M., Zitkus Z., Valius M. e-mail address bounced - despite being the one used to request curation. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt 14-Mar-2012: Contacted by JYOTI. 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Only protein-protein interactions human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ger M., Zitkus Z., Valius M. e-mail address bounced - despite being the one used to request curation. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt Accepted 2011-DEC-15 by JYOTI 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Cell. Signal. (0898-6568) 2011 Only protein-protein interactions imex curation Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. Cell. Signal. (0898-6568) 2011 imex curation Ger M., Zitkus Z., Valius M. e-mail address bounced - despite being the one used to request curation. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt 14-Mar-2012: Contacted by JYOTI. 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Ger M., Zitkus Z., Valius M. e-mail address bounced - despite being the one used to request curation. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt Accepted 2011-DEC-15 by JYOTI 14-Mar-2012: Contacted by JYOTI. 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Cell. Signal. (0898-6568) 2011 Only protein-protein interactions imex curation Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. Cell. Signal. (0898-6568) 2011 imex curation Ger M., Zitkus Z., Valius M. e-mail address bounced - despite being the one used to request curation. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt 14-Mar-2012: Contacted by JYOTI. 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Only protein-protein interactions in vitro In vitro ms of complexes mass spectrometry studies of complexes ms of complexes ms participant Identification by mass spectrometry ms participant Ger M., Zitkus Z., Valius M. e-mail address bounced - despite being the one used to request curation. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt Accepted 2011-DEC-15 by JYOTI 14-Mar-2012: Contacted by JYOTI. 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Cell. Signal. (0898-6568) 2011 Only protein-protein interactions imex curation Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. Cell. Signal. (0898-6568) 2011 imex curation Ger M., Zitkus Z., Valius M. e-mail address bounced - despite being the one used to request curation. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt 14-Mar-2012: Contacted by JYOTI. 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Only protein-protein interactions human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Ger M., Zitkus Z., Valius M. e-mail address bounced - despite being the one used to request curation. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt Accepted 2011-DEC-15 by JYOTI 14-Mar-2012: Contacted by JYOTI. 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Cell. Signal. (0898-6568) 2011 Only protein-protein interactions imex curation Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. Adaptor protein Nck1 interacts with p120 Ras GTPase-activating protein and regulates its activity. Cell. Signal. (0898-6568) 2011 imex curation Ger M., Zitkus Z., Valius M. e-mail address bounced - despite being the one used to request curation. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt 14-Mar-2012: Contacted by JYOTI. 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting western blot western blot Immuno blot Immuno blot Ger M., Zitkus Z., Valius M. e-mail address bounced - despite being the one used to request curation. RasGAP activity could not be captures as source of H-Ras could not be traced. marija.ger@bchi.vu.lt, zigmantas.zitkus@bchi.vu.lt, mindaugas.valius@bchi.vu.lt Accepted 2011-DEC-15 by JYOTI 14-Mar-2012: Contacted by JYOTI. 2011-11-01: Requested by Mindaugas Valius, Proteomics Centre, Vilnius University Institute of Biochemistry, Mokslininku st. 12, Vilnius LT-08662, Lithuania Cell. Signal. (0898-6568) 2011 Only protein-protein interactions imex curation rasa1_human Ras GTPase-activating protein 1 Ras p21 protein activator p120GAP RASA1 RASA GAP protein protein human Homo sapiens Human MMAAEAGSEEGGPVTAGAGGGGAAAGSSAYPAVCRVKIPAALPVAAAPYPGLVETGVAGTLGGGAALGSEFLGAGSVAGALGGAGLTGGGTAAGVAGAAAGVAGAAVAGPSGDMALTKLPTSLLAETLGPGGGFPPLPPPPYLPPLGAGLGTVDEGDSLDGPEYEEEEVAIPLTAPPTNQWYHGKLDRTIAEERLRQAGKSGSYLIRESDRRPGSFVLSFLSQMNVVNHFRIIAMCGDYYIGGRRFSSLSDLIGYYSHVSCLLKGEKLLYPVAPPEPVEDRRRVRAILPYTKVPDTDEISFLKGDMFIVHNELEDGWMWVTNLRTDEQGLIVEDLVEEVGREEDPHEGKIWFHGKISKQEAYNLLMTVGQVCSFLVRPSDNTPGDYSLYFRTNENIQRFKICPTPNNQFMMGGRYYNSIGDIIDHYRKEQIVEGYYLKEPVPMQDQEQVLNDTVDGKEIYNTIRRKTKDAFYKNIVKKGYLLKKGKGKRWKNLYFILEGSDAQLIYFESEKRATKPKGLIDLSVCSVYVVHDSLFGRPNCFQIVVQHFSEEHYIFYFAGETPEQAEDWMKGLQAFCNLRKSSPGTSNKRLRQVSSLVLHIEEAHKLPVKHFTNPYCNIYLNSVQVAKTHAREGQNPVWSEEFVFDDLPPDINRFEITLSNKTKKSKDPDILFMRCQLSRLQKGHATDEWFLLSSHIPLKGIEPGSLRVRARYSMEKIMPEEEYSEFKELILQKELHVVYALSHVCGQDRTLLASILLRIFLHEKLESLLLCTLNDREISMEDEATTLFRATTLASTLMEQYMKATATQFVHHALKDSILKIMESKQSCELSPSKLEKNEDVNTNLTHLLNILSELVEKIFMASEILPPTLRYIYGCLQKSVQHKWPTNTTMRTRVVSGFVFLRLICPAILNPRMFNIISDSPSPIAARTLILVAKSVQNLANLVEFGAKEPYMEGVNPFIKSNKHRMIMFLDELGNVPELPDTTEHSRTDLSRDLAALHEICVAHSDELRTLSNERGAQQHVLKKLLAITELLQQKQNQYTKTNDVR DIP protein P20936 original sequence version: 136 DIP protein P20936 original sequence version: 182 C35B6567F5BC5370 nck1_human Cytoplasmic protein NCK1 NCK1 NCK NCK adaptor protein 1 SH2/SH3 adaptor protein NCK-alpha protein protein human Homo sapiens Human MAEEVVVVAKFDYVAQQEQELDIKKNERLWLLDDSKSWWRVRNSMNKTGFVPSNYVERKNSARKASIVKNLKDTLGIGKVKRKPSVPDSASPADDSFVDPGERLYDLNMPAYVKFNYMAEREDELSLIKGTKVIVMEKCSDGWWRGSYNGQVGWFPSNYVTEEGDSPLGDHVGSLSEKLAAVVNNLNTGQVLHVVQALYPFSSSNDEELNFEKGDVMDVIEKPENDPEWWKCRKINGMVGLVPKNYVTVMQNNPLTSGLEPSPPQCDYIRPSLTGKFAGNPWYYGKVTRHQAEMALNERGHEGDFLIRDSESSPNDFSVSLKAQGKNKHFKVQLKETVYCIGQRKFSTMEELVEHYKKAPIFTSEQGEKLYLVKHLS 554E9B1A936AEF30 rasa1-nck1 1 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Fig. 1B nck1-rasa1 1 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Fig. 1B nck1-rasa1-1 4 7 unspecified role unspecified role bait bait P16333:p.Trp38Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain W K Wed May 02 18:37:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp38lys' Tue Jun 30 19:55:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp38Lys' P16333:p.[Trp38Lys;Trp143Lys;Trp229Lys;Arg308Lys] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain W K certain certain sequence position certain certain certain sequence position certain W K certain certain sequence position certain certain certain sequence position certain W K certain certain sequence position certain certain certain sequence position certain R K Wed May 02 15:06:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp38lys,trp143lys,trp229lys,arg308lys' Tue Jun 30 19:29:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp38Lys;Trp143Lys;Trp229Lys;Arg308Lys]' P16333:p.[Trp38Lys;Trp229Lys;Arg308Lys] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain W K certain certain sequence position certain certain certain sequence position certain R K certain certain sequence position certain certain certain sequence position certain W K Fri Jul 22 10:59:57 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'trp38lys,arg308lys,trp229lys' Wed May 02 18:01:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp38lys,trp229lys,arg308lys' Tue Jun 30 15:54:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp38Lys;Trp229Lys;Arg308Lys]' region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P16333:p.Trp229Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain W K Wed May 02 16:12:21 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp229lys' Tue Jun 30 18:12:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp229Lys' P16333:p.[Trp38_Trp39delinsLysLys;Trp143Lys;Trp229Lys] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain WW KK certain certain sequence position certain certain certain sequence position certain W K certain certain sequence position certain certain certain sequence position certain W K Wed May 02 15:59:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp_trp38-39lys_lys,trp143lys,trp229lys' Tue Jun 30 17:03:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp38_Trp39delinsLysLys;Trp143Lys;Trp229Lys]' P16333:p.[Trp143Lys;Trp229Lys;Arg308Lys] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain W K certain certain sequence position certain certain certain sequence position certain W K certain certain sequence position certain certain certain sequence position certain R K Wed May 02 15:17:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp143lys,trp229lys,arg308lys' Tue Jun 30 15:59:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp143Lys;Trp229Lys;Arg308Lys]' 6 unspecified role unspecified role prey prey physical association physical association Fig. 2A, 2B nck1-rasa1-2 2 7 unspecified role unspecified role bait bait region 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 sh3 region sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). 27 29 direct interaction direct interaction Fig. 2C nck1-rasa1-3 5 7 unspecified role unspecified role bait bait region 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 sh3 region sufficient to bind sufficient binding region sufficient to bind range range range range ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope 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 region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). direct interaction direct interaction Fig. 3A nck1-rasa1-4 3 7 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined human-hep_g2 Homo sapiens hepatocellular carcinoma (HCC) cells (p53-wt). hep_g2 human hepatocellular carcinoma (HCC) cells (p53-wt). direct interaction direct interaction Fig. 3C Molecular weight of complex = 51 474 Da.