MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it Ubiquitylation-dependent downregulation of Nck regulates its functional activity. Ubiquitylation-dependent downregulation of Nck regulates its functional activity. FEBS Lett. (0014-5793) 2014 imex curation Joseph N., Reicher B., David A., Matalon O., Barda-Saad M. Mira.Barda-Saad@biu.ac.il 20-Oct-2014: Contacted by IntAct-Help. Only protein-protein interactions human Homo sapiens Human anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Mira.Barda-Saad@biu.ac.il Accepted 2014-AUG-28 by LUANA 2014 imex curation 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 cblb_human E3 ubiquitin-protein ligase CBL-B Signal transduction protein CBL-B SH3-binding protein CBL-B Casitas B-lineage lymphoma proto-oncogene b RING finger protein 56 CBLB RNF56 Nbla00127 RING-type E3 ubiquitin transferase CBL-B protein protein human Homo sapiens Human MANSMNGRNPGGRGGNPRKGRILGIIDAIQDAVGPPKQAAADRRTVEKTWKLMDKVVRLCQNPKLQLKNSPPYILDILPDTYQHLRLILSKYDDNQKLAQLSENEYFKIYIDSLMKKSKRAIRLFKEGKERMYEEQSQDRRNLTKLSLIFSHMLAEIKAIFPNGQFQGDNFRITKADAAEFWRKFFGDKTIVPWKVFRQCLHEVHQISSGLEAMALKSTIDLTCNDYISVFEFDIFTRLFQPWGSILRNWNFLAVTHPGYMAFLTYDEVKARLQKYSTKPGSYIFRLSCTRLGQWAIGYVTGDGNILQTIPHNKPLFQALIDGSREGFYLYPDGRSYNPDLTGLCEPTPHDHIKVTQEQYELYCEMGSTFQLCKICAENDKDVKIEPCGHLMCTSCLTAWQESDGQGCPFCRCEIKGTEPIIVDPFDPRDEGSRCCSIIDPFGMPMLDLDDDDDREESLMMNRLANVRKCTDRQNSPVTSPGSSPLAQRRKPQPDPLQIPHLSLPPVPPRLDLIQKGIVRSPCGSPTGSPKSSPCMVRKQDKPLPAPPPPLRDPPPPPPERPPPIPPDNRLSRHIHHVESVPSRDPPMPLEAWCPRDVFGTNQLVGCRLLGEGSPKPGITASSNVNGRHSRVGSDPVLMRKHRRHDLPLEGAKVFSNGHLGSEEYDVPPRLSPPPPVTTLLPSIKCTGPLANSLSEKTRDPVEEDDDEYKIPSSHPVSLNSQPSHCHNVKPPVRSCDNGHCMLNGTHGPSSEKKSNIPDLSIYLKGDVFDSASDPVPLPPARPPTRDNPKHGSSLNRTPSDYDLLIPPLGEDAFDALPPSLPPPPPPARHSLIEHSKPPGSSSRPSSGQDLFLLPSDPFVDLASGQVPLPPARRLPGENVKTNRTSQDYDQLPSCSDGSQAPARPPKPRPRRTAPEIHHRKPHGPEAALENVDAKIAKLMGEGYAFEEVKRALEIAQNNVEVARSILREFAFPPPVSPRLNL B4E307D31B9F0779 cblb-nck1 1 Nck 2 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell P16333:p.Trp38Lys mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain W K Wed May 02 20:29:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp38lys' Tue Jun 30 19:20:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp38Lys' yfp tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Cbl-b 3 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell association association f6b f6c f6d 293T cells were co-transfected with YFP-tagged Nck constructs encoding Nck-wt, or each of the SH3 mutant molecules, together with HA-Ub and Cbl-b. As shown in Figure 6B, each of the Nck-SH3-mutants exhibited a substantial decrease in their 11 interaction with Cbl-b, and in Nck ubiquitylation. Moreover, a triple Nck-SH3 mutant (3SH3*) did not interact with Cbl-b at all, and was substantially less ubiquitylated (Fig. 6C). nck1-cblb-3 1 Cbl-b 3 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell Nck 2 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell yfp tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P16333:p.Trp143Lys mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain W K Wed May 02 19:35:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp143lys' Tue Jun 30 16:24:49 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp143Lys' association association f6b f6c f6d 293T cells were co-transfected with YFP-tagged Nck constructs encoding Nck-wt, or each of the SH3 mutant molecules, together with HA-Ub and Cbl-b. As shown in Figure 6B, each of the Nck-SH3-mutants exhibited a substantial decrease in their 11 interaction with Cbl-b, and in Nck ubiquitylation. Moreover, a triple Nck-SH3 mutant (3SH3*) did not interact with Cbl-b at all, and was substantially less ubiquitylated (Fig. 6C). nck1-cblb-4 1 Nck 2 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell yfp tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P16333:p.Trp229Lys mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain W K Wed May 02 19:09:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp229lys' Tue Jun 30 17:59:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp229Lys' Cbl-b 3 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell association association f6b f6c f6d 293T cells were co-transfected with YFP-tagged Nck constructs encoding Nck-wt, or each of the SH3 mutant molecules, together with HA-Ub and Cbl-b. As shown in Figure 6B, each of the Nck-SH3-mutants exhibited a substantial decrease in their 11 interaction with Cbl-b, and in Nck ubiquitylation. Moreover, a triple Nck-SH3 mutant (3SH3*) did not interact with Cbl-b at all, and was substantially less ubiquitylated (Fig. 6C). nck1-cblb-5 1 Cbl-b 3 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell Nck 2 western blot western blot Immuno blot Immuno blot western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell P16333:p.[Trp38Lys;Trp143Lys;Trp229Lys] mutation disrupting mutation disrupting interaction mutation disrupting 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 Fri Jul 22 11:09:24 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'trp229lys,trp38lys,trp143lys' Wed May 02 17:04:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp38lys,trp143lys,trp229lys' Tue Jun 30 15:48:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Trp38Lys;Trp143Lys;Trp229Lys]' yfp tag yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association f6b f6c f6d 293T cells were co-transfected with YFP-tagged Nck constructs encoding Nck-wt, or each of the SH3 mutant molecules, together with HA-Ub and Cbl-b. As shown in Figure 6B, each of the Nck-SH3-mutants exhibited a substantial decrease in their interaction with Cbl-b, and in Nck ubiquitylation. Moreover, a triple Nck-SH3 mutant (3SH3*) did not interact with Cbl-b at all, and was substantially less ubiquitylated (Fig. 6C). To confirm this observation in T cells, Jurkat T-cell lines stably expressing the YFPNck constructs were prepared and analyzed (Fig. 6D). As in 293T cells, the molecular association of Cbl-b with each individual Nck-SH3-mutant was abolished. Furthermore, Nck ubiquitylation was correspondingly decreased. Thus, the three SH3 domains of Nck are all required for stable interaction with Cbl-b, and consequently, for Nck ubiquitylation.