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/ Efficient ADAM22 surface expression is mediated by phosphorylation-dependent interaction with 14-3-3 protein family members. Efficient ADAM22 surface expression is mediated by phosphorylation-dependent interaction with 14-3-3 protein family members. J. Cell. Sci. (0021-9533) 2006 imex curation Godde NJ., D'Abaco GM., Paradiso L., Novak U. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer u.novak@unimelb.edu.au 22-Jan-2008: Contacted by JYOTI. Human brain cDNA davet 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. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Godde NJ., D'Abaco GM., Paradiso L., Novak U. Synapse - Interactions of proteins with an established role in the presynapse. The source of proteins 14-3-3 zeta, gamma and tau were taken from PMIDs 7615088, 12507503 and 10840038, respectively. u.novak@unimelb.edu.au Accepted 2007-Not a month!-19 by DAVET. 22-Jan-2008: Contacted by JYOTI. Human brain cDNA J. Cell. Sci. (0021-9533) 2006 davet Only protein-protein interactions imex curation Efficient ADAM22 surface expression is mediated by phosphorylation-dependent interaction with 14-3-3 protein family members. Efficient ADAM22 surface expression is mediated by phosphorylation-dependent interaction with 14-3-3 protein family members. J. Cell. Sci. (0021-9533) 2006 imex curation Godde NJ., D'Abaco GM., Paradiso L., Novak U. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer u.novak@unimelb.edu.au 22-Jan-2008: Contacted by JYOTI. Human brain cDNA davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Godde NJ., D'Abaco GM., Paradiso L., Novak U. Synapse - Interactions of proteins with an established role in the presynapse. u.novak@unimelb.edu.au Accepted 2007-Not a month!-19 by DAVET. Human brain cDNA J. Cell. Sci. (0021-9533) 2006 davet Only protein-protein interactions imex curation Efficient ADAM22 surface expression is mediated by phosphorylation-dependent interaction with 14-3-3 protein family members. Efficient ADAM22 surface expression is mediated by phosphorylation-dependent interaction with 14-3-3 protein family members. J. Cell. Sci. (0021-9533) 2006 imex curation Godde NJ., D'Abaco GM., Paradiso L., Novak U. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer u.novak@unimelb.edu.au 22-Jan-2008: Contacted by JYOTI. Human brain cDNA davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Godde NJ., D'Abaco GM., Paradiso L., Novak U. Synapse - Interactions of proteins with an established role in the presynapse. u.novak@unimelb.edu.au Accepted 2007-Not a month!-19 by DAVET. 22-Jan-2008: Contacted by JYOTI. Human brain cDNA J. Cell. Sci. (0021-9533) 2006 davet Only protein-protein interactions imex curation Efficient ADAM22 surface expression is mediated by phosphorylation-dependent interaction with 14-3-3 protein family members. Efficient ADAM22 surface expression is mediated by phosphorylation-dependent interaction with 14-3-3 protein family members. J. Cell. Sci. (0021-9533) 2006 imex curation Godde NJ., D'Abaco GM., Paradiso L., Novak U. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer u.novak@unimelb.edu.au 22-Jan-2008: Contacted by JYOTI. Human brain cDNA davet Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Godde NJ., D'Abaco GM., Paradiso L., Novak U. Synapse - Interactions of proteins with an established role in the presynapse. u.novak@unimelb.edu.au Accepted 2007-Not a month!-19 by DAVET. 22-Jan-2008: Contacted by JYOTI. Human brain cDNA J. Cell. Sci. (0021-9533) 2006 davet Only protein-protein interactions imex curation q9p0k1-3 Disintegrin and metalloproteinase domain-containing protein 22 ADAM22 MDC2 Metalloproteinase-disintegrin ADAM22-3 Metalloproteinase-like, disintegrin-like, and cysteine-rich protein 2 Alpha protein protein human Homo sapiens Human MQAAVAVSVPFLLLCVLGTCPPARCGQAGDASLMELEKRKENRFVERQSIVPLRLIYRSGGEDESRHDALDTRVRGDLGGPQLTHVDQASFQVDAFGTSFILDVVLNHDLLSSEYIERHIEHGGKTVEVKGGEHCYYQGHIRGNPDSFVALSTCHGLHGMFYDGNHTYLIEPEENDTTQEDFHFHSVYKSRLFEFSLDDLPSEFQQVNITPSKFILKPRPKRSKRQLRRYPRNVEEETKYIELMIVNDHLMFKKHRLSVVHTNTYAKSVVNMADLIYKDQLKTRIVLVAMETWATDNKFAISENPLITLREFMKYRRDFIKEKSDAVHLFSGSQFESSRSGAAYIGGICSLLKGGGVNEFGKTDLMAVTLAQSLAHNIGIISDKRKLASGECKCEDTWSGCIMGDTGYYLPKKFTQCNIEEYHDFLNSGGGACLFNKPSKLLDPPECGNGFIETGEECDCGTPAECVLEGAECCKKCTLTQDSQCSDGLCCKKCKFQPMGTVCREAVNDCDIRETCSGNSSQCAPNIHKMDGYSCDGVQGICFGGRCKTRDRQCKYIWGQKVTASDKYCYEKLNIEGTEKGNCGKDKDTWIQCNKRDVLCGYLLCTNIGNIPRLGELDGEITSTLVVQQGRTLNCSGGHVKLEEDVDLGYVEDGTPCGPQMMCLEHRCLPVASFNFSTCLSSKEGTICSGNGVCSNELKCVCNRHWIGSDCNTYFPHNDDAKTGITLSGNGVAGTNIIIGIIAGTILVLALILGITAWGYKNYREQRQLPQGDYVKKPGDGDSFYSDIPPGVSTNSASSSKKRSNGLSHSWSERIPDTKHISDICENGRPRSNSWQGNLGGNKKKIRGKRFRPRSNSTE 6105B681A1331D55 1433e_human 14-3-3 protein epsilon YWHAE protein protein human Homo sapiens Human MDDREDLVYQAKLAEQAERYDEMVESMKKVAGMDVELTVEERNLLSVAYKNVIGARRASWRIISSIEQKEENKGGEDKLKMIREYRQMVETELKLICCDILDVLDKHLIPAANTGESKVFYYKMKGDYHRYLAEFATGNDRKEAAENSLVAYKAASDIAMTELPPTHPIRLGLALNFSVFYYEILNSPDRACRLAKAAFDDAIAELDTLSEESYKDSTLIMQLLRDNLTLWTSDMQGDGEEQNKEALQDVEDENQ DIP protein P62258 original sequence version: 130 DIP protein P62258 original sequence version: 132 DIP protein P62258 original sequence version: 91 07817CCBD1F75B26 1433g_mouse 14-3-3 protein gamma Ywhag protein protein mouse Mus musculus Mouse MVDREQLVQKARLAEQAERYDDMAAAMKNVTELNEPLSNEERNLLSVAYKNVVGARRSSWRVISSIEQKTSADGNEKKIEMVRAYREKIEKELEAVCQDVLSLLDNYLIKNCSETQYESKVFYLKMKGDYYRYLAEVATGEKRATVVESSEKAYSEAHEISKEHMQPTHPIRLGLALNYSVFYYEIQNAPEQACHLAKTAFDDAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDQQDDDGGEGNN B0D16C6DE1F4455D 1433f_human 14-3-3 protein eta YWHAH YWHA1 Protein AS1 protein protein human Homo sapiens Human MGDREQLLQRARLAEQAERYDDMASAMKAVTELNEPLSNEDRNLLSVAYKNVVGARRSSWRVISSIEQKTMADGNEKKLEKVKAYREKIEKELETVCNDVLSLLDKFLIKNCNDFQYESKVFYLKMKGDYYRYLAEVASGEKKNSVVEASEAAYKEAFEISKEQMQPTHPIRLGLALNFSVFYYEIQNAPEQACLLAKQAFDDAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDQQDEEAGEGN DIP protein Q04917 original sequence version: 166 DIP protein Q04917 original sequence version: 168 DIP protein Q04917 original sequence version: 175 D70FBC100C45D6E5 1433b_human 14-3-3 protein beta/alpha YWHAB Protein kinase C inhibitor protein 1 Protein 1054 protein protein human Homo sapiens Human MTMDKSELVQKAKLAEQAERYDDMAAAMKAVTEQGHELSNEERNLLSVAYKNVVGARRSSWRVISSIEQKTERNEKKQQMGKEYREKIEAELQDICNDVLELLDKYLIPNATQPESKVFYLKMKGDYFRYLSEVASGDNKQTTVSNSQQAYQEAFEISKKEMQPTHPIRLGLALNFSVFYYEILNSPEKACSLAKTAFDEAIAELDTLNEESYKDSTLIMQLLRDNLTLWTSENQGDEGDAGEGEN DIP protein P31946 original sequence version: 171 DIP protein P31946 original sequence version: 97 DIP protein P31946 original sequence version: 119 DIP protein P31946 original sequence version: 122 DIP protein P31946 original sequence version: 180 6BE1A9BF97468017 1433z_human 14-3-3 protein zeta/delta YWHAZ Protein kinase C inhibitor protein 1 protein protein human Homo sapiens Human MDKNELVQKAKLAEQAERYDDMAACMKSVTEQGAELSNEERNLLSVAYKNVVGARRSSWRVVSSIEQKTEGAEKKQQMAREYREKIETELRDICNDVLSLLEKFLIPNASQAESKVFYLKMKGDYYRYLAEVAAGDDKKGIVDQSQQAYQEAFEISKKEMQPTHPIRLGLALNFSVFYYEILNSPEKACSLAKTAFDEAIAELDTLSEESYKDSTLIMQLLRDNLTLWTSDTQGDEAEAGEGGEN DIP protein P63104 original sequence version: 144 D464DF2286BBFE60 1433t_human 14-3-3 protein theta YWHAQ 14-3-3 protein tau 14-3-3 protein T-cell Protein HS1 protein protein human Homo sapiens Human MEKTELIQKAKLAEQAERYDDMATCMKAVTEQGAELSNEERNLLSVAYKNVVGGRRSAWRVISSIEQKTDTSDKKLQLIKDYREKVESELRSICTTVLELLDKYLIANATNPESKVFYLKMKGDYFRYLAEVACGDDRKQTIDNSQGAYQEAFDISKKEMQPTHPIRLGLALNFSVFYYEILNNPELACTLAKTAFDEAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDSAGEECDAAEGAEN DIP protein P27348 original sequence version: 164 DIP protein P27348 original sequence version: 173 175534325E9E37C4 q9p0k1-1 Disintegrin and metalloproteinase domain-containing protein 22 ADAM22 MDC2 Metalloproteinase-disintegrin ADAM22-3 Metalloproteinase-like, disintegrin-like, and cysteine-rich protein 2 Epsilon protein protein human Homo sapiens Human MQAAVAVSVPFLLLCVLGTCPPARCGQAGDASLMELEKRKENRFVERQSIVPLRLIYRSGGEDESRHDALDTRVRGDLGGPQLTHVDQASFQVDAFGTSFILDVVLNHDLLSSEYIERHIEHGGKTVEVKGGEHCYYQGHIRGNPDSFVALSTCHGLHGMFYDGNHTYLIEPEENDTTQEDFHFHSVYKSRLFEFSLDDLPSEFQQVNITPSKFILKPRPKRSKRQLRRYPRNVEEETKYIELMIVNDHLMFKKHRLSVVHTNTYAKSVVNMADLIYKDQLKTRIVLVAMETWATDNKFAISENPLITLREFMKYRRDFIKEKSDAVHLFSGSQFESSRSGAAYIGGICSLLKGGGVNEFGKTDLMAVTLAQSLAHNIGIISDKRKLASGECKCEDTWSGCIMGDTGYYLPKKFTQCNIEEYHDFLNSGGGACLFNKPSKLLDPPECGNGFIETGEECDCGTPAECVLEGAECCKKCTLTQDSQCSDGLCCKKCKFQPMGTVCREAVNDCDIRETCSGNSSQCAPNIHKMDGYSCDGVQGICFGGRCKTRDRQCKYIWGQKVTASDKYCYEKLNIEGTEKGNCGKDKDTWIQCNKRDVLCGYLLCTNIGNIPRLGELDGEITSTLVVQQGRTLNCSGGHVKLEEDVDLGYVEDGTPCGPQMMCLEHRCLPVASFNFSTCLSSKEGTICSGNGVCSNELKCVCNRHWIGSDCNTYFPHNDDAKTGITLSGNGVAGTNIIIGIIAGTILVLALILGITAWGYKNYREQRQLPQGDYVKKPGDGDSFYSDIPPGVSTNSASSSKKRSNGLSHSWSERIPDTKHISDICENGRPRSNSWQGNLGGNKKKIRGKRFRPRSNSTETLSPAKSPSSSTGSIASSRKYPYPMPPLPDEDKKVNRQSARLWETSI 265ECCD0FA6C088B adam22-ywhab 2 9 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait c-teminal cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association Sequential transformations of pDBLeu-ADAM22v1 cytoplasmic domain and a human brain cDNA library into yeast cells resulted in an estimated 6.2 106 transformants, which were subsequently screened for His3 reporter gene activation. The 23 clones deemed positive for His3 selection were then subjected to second round screening for lacZ reporter activation by X-gal assay, resulting in 13 positive clones. Sequencing of yeast plasmid DNA recovered in E. coli revealed the identity of 10 clones. All but one of these belonged to the 14-3-3 protein family, including isoforms beta(), zeta ()and tau () adam22-ywhaz 2 12 unspecified role unspecified role bait bait c-teminal cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 10 unspecified role unspecified role prey prey physical association physical association Sequential transformations of pDBLeu-ADAM22v1 cytoplasmic domain and a human brain cDNA library into yeast cells resulted in an estimated 6.2 106 transformants, which were subsequently screened for His3 reporter gene activation. The 23 clones deemed positive for His3 selection were then subjected to second round screening for lacZ reporter activation by X-gal assay, resulting in 13 positive clones. Sequencing of yeast plasmid DNA recovered in E. coli revealed the identity of 10 clones. All but one of these belonged to the 14-3-3 protein family, including isoforms beta(), zeta ()and tau () adam22-ywhaq 2 12 unspecified role unspecified role bait bait c-teminal cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 11 unspecified role unspecified role prey prey physical association physical association Sequential transformations of pDBLeu-ADAM22v1 cytoplasmic domain and a human brain cDNA library into yeast cells resulted in an estimated 6.2 106 transformants, which were subsequently screened for His3 reporter gene activation. The 23 clones deemed positive for His3 selection were then subjected to second round screening for lacZ reporter activation by X-gal assay, resulting in 13 positive clones. Sequencing of yeast plasmid DNA recovered in E. coli revealed the identity of 10 clones. All but one of these belonged to the 14-3-3 protein family, including isoforms beta(), zeta ()and tau () adam22-ywhaz-1 3 12 unspecified role unspecified role bait bait c-teminal cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Q9P0K1-1:p.[Arg831Gln;Ser832Gly;Arg854Gln;Ser855Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S G certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S A Wed May 02 19:18:56 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg831gln,ser832gly,arg854gln,ser855ala' Tue Jun 30 18:11:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg831Gln;Ser832Gly;Arg854Gln;Ser855Ala]' 10 unspecified role unspecified role prey prey physical association physical association All but one of these belonged to the 14-3-3 protein family, including isoforms beta (), zeta () and tau (). The remaining clone consisted of unidentifiable genomic sequence. Recovered 14-3-3 plasmid DNA or empty library vector pPC86 were re-introduced into yeast with either pDBLeu, pDBLeu-ADAM22v1 cytoplasmic domain or pDBLeu- ADAM22v4 cytoplasmic domain bait constructs. These yeast transformants were then assayed against the three reporter genes His3, lacZ and URA3 to confirm interactions in the yeast two-hybrid system 2, 4 C adam22-ywhab-1 3 9 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait Q9P0K1-1:p.[Arg831Gln;Ser832Gly;Arg854Gln;Ser855Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S G certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S A Wed May 02 19:38:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg831gln,ser832gly,arg854gln,ser855ala' Tue Jun 30 20:05:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg831Gln;Ser832Gly;Arg854Gln;Ser855Ala]' c-teminal cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association All but one of these belonged to the 14-3-3 protein family, including isoforms beta (), zeta () and tau (). The remaining clone consisted of unidentifiable genomic sequence. Recovered 14-3-3 plasmid DNA or empty library vector pPC86 were re-introduced into yeast with either pDBLeu, pDBLeu-ADAM22v1 cytoplasmic domain or pDBLeu- ADAM22v4 cytoplasmic domain bait constructs. These yeast transformants were then assayed against the three reporter genes His3, lacZ and URA3 to confirm interactions in the yeast two-hybrid system 2, 4 C adam22-ywhaq-1 3 11 unspecified role unspecified role prey prey 12 unspecified role unspecified role bait bait Q9P0K1-1:p.[Arg831Gln;Ser832Gly;Arg854Gln;Ser855Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S G certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S A Wed May 02 17:37:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg831gln,ser832gly,arg854gln,ser855ala' Tue Jun 30 19:16:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg831Gln;Ser832Gly;Arg854Gln;Ser855Ala]' c-teminal cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association All but one of these belonged to the 14-3-3 protein family, including isoforms beta (), zeta () and tau (). The remaining clone consisted of unidentifiable genomic sequence. Recovered 14-3-3 plasmid DNA or empty library vector pPC86 were re-introduced into yeast with either pDBLeu, pDBLeu-ADAM22v1 cytoplasmic domain or pDBLeu- ADAM22v4 cytoplasmic domain bait constructs. These yeast transformants were then assayed against the three reporter genes His3, lacZ and URA3 to confirm interactions in the yeast two-hybrid system 2, 4 C adam22-ywhaq-2 3 11 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1567398]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 860-860, sequence length 859 [EBI-1567398]760-860 physical association physical association All but one of these belonged to the 14-3-3 protein family, including isoforms beta (), zeta () and tau (). The remaining clone consisted of unidentifiable genomic sequence. Recovered 14-3-3 plasmid DNA or empty library vector pPC86 were re-introduced into yeast with either pDBLeu, pDBLeu-ADAM22v1 cytoplasmic domain or pDBLeu- ADAM22v4 cytoplasmic domain bait constructs. These yeast transformants were then assayed against the three reporter genes His3, lacZ and URA3 to confirm interactions in the yeast two-hybrid system 2 adam22-ywhaz-2 3 10 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1567405]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 860-860, sequence length 859 [EBI-1567405]760-860 physical association physical association All but one of these belonged to the 14-3-3 protein family, including isoforms beta (), zeta () and tau (). The remaining clone consisted of unidentifiable genomic sequence. Recovered 14-3-3 plasmid DNA or empty library vector pPC86 were re-introduced into yeast with either pDBLeu, pDBLeu-ADAM22v1 cytoplasmic domain or pDBLeu- ADAM22v4 cytoplasmic domain bait constructs. These yeast transformants were then assayed against the three reporter genes His3, lacZ and URA3 to confirm interactions in the yeast two-hybrid system 2 adam22-ywhab-2 3 5 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1567413]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 860-860, sequence length 859 [EBI-1567413]760-860 9 unspecified role unspecified role prey prey physical association physical association All but one of these belonged to the 14-3-3 protein family, including isoforms beta (), zeta () and tau (). The remaining clone consisted of unidentifiable genomic sequence. Recovered 14-3-3 plasmid DNA or empty library vector pPC86 were re-introduced into yeast with either pDBLeu, pDBLeu-ADAM22v1 cytoplasmic domain or pDBLeu- ADAM22v4 cytoplasmic domain bait constructs. These yeast transformants were then assayed against the three reporter genes His3, lacZ and URA3 to confirm interactions in the yeast two-hybrid system 2, 4 C ywhab-adam22 1 5 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 Q9P0K1-3:p.[Arg854Gln;Ser855Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S A Wed May 02 19:37:09 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg854gln,ser855ala' Tue Jun 30 16:18:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg854Gln;Ser855Ala]' Q9P0K1-3:p.[Arg831Gln;Ser832Gly] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S G Wed May 02 15:19:13 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg831gln,ser832gly' Tue Jun 30 18:55:33 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg831Gln;Ser832Gly]' Q9P0K1-3:p.[Arg831Gln;Ser832Gly;Arg854Gln;Ser855Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S G certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S A Wed May 02 19:39:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg831gln,ser832gly,arg854gln,ser855ala' Tue Jun 30 20:17:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg831Gln;Ser832Gly;Arg854Gln;Ser855Ala]' c-terminal required to bind necessary binding region required to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1567529]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 906-906, sequence length 859 [EBI-1567529]831-906 9 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association ADAM22 interacts with all 14-3-3 protein family members expressed in the brain. HEK 293T cells co-expressing FLAG-tagged full length ADAM22v4 and Myc-tagged full length 14-3-3 proteins , , , , or HA-tagged 14-3-3 were lysed and proteins were immunoprecipitated (IP) with anti-Myc or anti-HA antibodies and probed with anti-FLAG antibody. Immunoblots were then re-probed with anti-Myc or anti-HA antibodies. 3, 4 A, 5 A Q9P0K1-3; P31946 transfection Dephosphorylation of ADAM22 significantly reduced binding of all 14-3-3 isoforms. By contrast, dephosphorylation of 14-3-3 proteins alone did not impact on their ability to co-immunoprecipitate ADAM22. ywhag-adam22 1 5 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 7 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association ADAM22 interacts with all 14-3-3 protein family members expressed in the brain. HEK 293T cells co-expressing FLAG-tagged full length ADAM22v4 and Myc-tagged full length 14-3-3 proteins , , , , or HA-tagged 14-3-3 were lysed and proteins were immunoprecipitated (IP) with anti-Myc or anti-HA antibodies and probed with anti-FLAG antibody. Immunoblots were then re-probed with anti-Myc or anti-HA antibodies. 3, 5 A Q9P0K1-3; P61982 transfection Dephosphorylation of ADAM22 significantly reduced binding of all 14-3-3 isoforms. By contrast, dephosphorylation of 14-3-3 proteins alone did not impact on their ability to co-immunoprecipitate ADAM22. ywhaf-adam22 1 5 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 8 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association ADAM22 interacts with all 14-3-3 protein family members expressed in the brain. HEK 293T cells co-expressing FLAG-tagged full length ADAM22v4 and Myc-tagged full length 14-3-3 proteins , , , , or HA-tagged 14-3-3 were lysed and proteins were immunoprecipitated (IP) with anti-Myc or anti-HA antibodies and probed with anti-FLAG antibody. Immunoblots were then re-probed with anti-Myc or anti-HA antibodies. 3, 5 A Q9P0K1-3; Q04917 transfection Dephosphorylation of ADAM22 significantly reduced binding of all 14-3-3 isoforms. By contrast, dephosphorylation of 14-3-3 proteins alone did not impact on their ability to co-immunoprecipitate ADAM22. ywhae-adam22 1 5 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 6 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association ADAM22 interacts with all 14-3-3 protein family members expressed in the brain. HEK 293T cells co-expressing FLAG-tagged full length ADAM22v4 and Myc-tagged full length 14-3-3 proteins , , , , or HA-tagged 14-3-3 were lysed and proteins were immunoprecipitated (IP) with anti-Myc or anti-HA antibodies and probed with anti-FLAG antibody. Immunoblots were then re-probed with anti-Myc or anti-HA antibodies. 3, 5 A Q9P0K1-3; P62258 transfection Dephosphorylation of ADAM22 significantly reduced binding of all 14-3-3 isoforms. By contrast, dephosphorylation of 14-3-3 proteins alone did not impact on their ability to co-immunoprecipitate ADAM22. ywhaq-adam22 1 11 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 5 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 ADAM22 interacts with all 14-3-3 protein family members expressed in the brain. HEK 293T cells co-expressing FLAG-tagged full length ADAM22v4 and Myc-tagged full length 14-3-3 proteins , , , , or HA-tagged 14-3-3 were lysed and proteins were immunoprecipitated (IP) with anti-Myc or anti-HA antibodies and probed with anti-FLAG antibody. Immunoblots were then re-probed with anti-Myc or anti-HA antibodies. 3, 5 A Q9P0K1-3; P27348 transfection Dephosphorylation of ADAM22 significantly reduced binding of all 14-3-3 isoforms. By contrast, dephosphorylation of 14-3-3 proteins alone did not impact on their ability to co-immunoprecipitate ADAM22. ywhaz-adam22 1 5 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 10 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association ADAM22 interacts with all 14-3-3 protein family members expressed in the brain. HEK 293T cells co-expressing FLAG-tagged full length ADAM22v4 and Myc-tagged full length 14-3-3 proteins , , , , or HA-tagged 14-3-3 were lysed and proteins were immunoprecipitated (IP) with anti-Myc or anti-HA antibodies and probed with anti-FLAG antibody. Immunoblots were then re-probed with anti-Myc or anti-HA antibodies. 3, 5 A Q9P0K1-3; P63104 transfection Dephosphorylation of ADAM22 significantly reduced binding of all 14-3-3 isoforms. By contrast, dephosphorylation of 14-3-3 proteins alone did not impact on their ability to co-immunoprecipitate ADAM22. ywhaz-adam22-1 4 5 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 Q9P0K1-3:p.[Arg831Gln;Ser832Gly;Arg854Gln;Ser855Ala] mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S G certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S A Wed May 02 15:19:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg831gln,ser832gly,arg854gln,ser855ala' Tue Jun 30 19:11:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg831Gln;Ser832Gly;Arg854Gln;Ser855Ala]' Q9P0K1-3:p.[Arg831Gln;Ser832Gly] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R Q certain certain sequence position certain certain certain sequence position certain S G Wed May 02 15:41:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg831gln,ser832gly' Tue Jun 30 20:17:55 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg831Gln;Ser832Gly]' c-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1567527]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 906-906, sequence length 859 [EBI-1567527]831-906 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. 10 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 ecobb Escherichia coli (strain B / BL21) physical association physical association we performed GST-14-3-3 fusion protein pull-down assays using HEK 293T cell lysates expressing the same ADAM22 wild-type and mutant proteins (Fig. 4B). Mutations of the first binding site similarly reduced the ability of GST-14-3-3 to pull down the ADAM22 precursor form. Mutation of the second site had very little effect, whereas double mutation or deletion of both binding sites abolished binding to the GST-14-3-3 protein. 4 B