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/ ruusala-2008-3 The atypical Rho GTPase Wrch1 collaborates with the nonreceptor tyrosine kinases Pyk2 and Src in regulating cytoskeletal dynamics. 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 western blot western blot Immuno blot Immuno blot Ruusala A., Aspenstrom P. Apoptosis - Interactions involving proteins with a function related to apoptosis GAL4DB Wrch1(Q107L)-expressing yeast was used in a screen of a cDNA library from EBV-transformed human B cells fused to the GAL4 activation domain in the pACT vector. pontus.aspenstrom@licr.uu.se A cDNA library from EBV-transformed human B cells was used. Mol. Cell. Biol. (0270-7306) 2008 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer ruusala-2008-5 The atypical Rho GTPase Wrch1 collaborates with the nonreceptor tyrosine kinases Pyk2 and Src in regulating cytoskeletal dynamics. 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 Ruusala A., Aspenstrom P. Apoptosis - Interactions involving proteins with a function related to apoptosis GAL4DB Wrch1(Q107L)-expressing yeast was used in a screen of a cDNA library from EBV-transformed human B cells fused to the GAL4 activation domain in the pACT vector. pontus.aspenstrom@licr.uu.se A cDNA library from EBV-transformed human B cells was used. Mol. Cell. Biol. (0270-7306) 2008 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer ruusala-2008-1 The atypical Rho GTPase Wrch1 collaborates with the nonreceptor tyrosine kinases Pyk2 and Src in regulating cytoskeletal dynamics. 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 Ruusala A., Aspenstrom P. Apoptosis - Interactions involving proteins with a function related to apoptosis GAL4DB Wrch1(Q107L)-expressing yeast was used in a screen of a cDNA library from EBV-transformed human B cells fused to the GAL4 activation domain in the pACT vector. pontus.aspenstrom@licr.uu.se A cDNA library from EBV-transformed human B cells was used. Mol. Cell. Biol. (0270-7306) 2008 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer ruusala-2008-2 The atypical Rho GTPase Wrch1 collaborates with the nonreceptor tyrosine kinases Pyk2 and Src in regulating cytoskeletal dynamics. 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 Ruusala A., Aspenstrom P. Apoptosis - Interactions involving proteins with a function related to apoptosis GAL4DB Wrch1(Q107L)-expressing yeast was used in a screen of a cDNA library from EBV-transformed human B cells fused to the GAL4 activation domain in the pACT vector. pontus.aspenstrom@licr.uu.se A cDNA library from EBV-transformed human B cells was used. Mol. Cell. Biol. (0270-7306) 2008 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer ruusala-2008-4 The atypical Rho GTPase Wrch1 collaborates with the nonreceptor tyrosine kinases Pyk2 and Src in regulating cytoskeletal dynamics. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Ruusala A., Aspenstrom P. Apoptosis - Interactions involving proteins with a function related to apoptosis GAL4DB Wrch1(Q107L)-expressing yeast was used in a screen of a cDNA library from EBV-transformed human B cells fused to the GAL4 activation domain in the pACT vector. pontus.aspenstrom@licr.uu.se A cDNA library from EBV-transformed human B cells was used. Mol. Cell. Biol. (0270-7306) 2008 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer rhou_human Rho-related GTP-binding protein RhoU RHOU ARHU CDC42L1 G28K WRCH1 SB128 Wnt-1 responsive Cdc42 homolog 1 Ryu GTPase CDC42-like GTPase 1 Rho GTPase-like protein ARHU GTP-binding protein-like 1 protein protein human Homo sapiens Human MPPQQGDPAFPDRCEAPPVPPRRERGGRGGRGPGEPGGRGRAGGAEGRGVKCVLVGDGAVGKTSLVVSYTTNGYPTEYIPTAFDNFSAVVSVDGRPVRLQLCDTAGQDEFDKLRPLCYTNTDIFLLCFSVVSPSSFQNVSEKWVPEIRCHCPKAPIILVGTQSDLREDVKVLIELDKCKEKPVPEEAAKLCAEEIKAASYIECSALTQKNLKEVFDAAIVAGIQYSDTQQQPKKSKSRTPDKMKNLSKSWWKKYCCFV fak2_human Protein-tyrosine kinase 2-beta PTK2B FAK2 PYK2 RAFTK Focal adhesion kinase 2 Proline-rich tyrosine kinase 2 Cell adhesion kinase beta Calcium-dependent tyrosine kinase Related adhesion focal tyrosine kinase Calcium-regulated non-receptor proline-rich tyrosine kinase protein protein human Homo sapiens Human MSGVSEPLSRVKLGTLRRPEGPAEPMVVVPVDVEKEDVRILKVCFYSNSFNPGKNFKLVKCTVQTEIREIITSILLSGRIGPNIRLAECYGLRLKHMKSDEIHWLHPQMTVGEVQDKYECLHVEAEWRYDLQIRYLPEDFMESLKEDRTTLLYFYQQLRNDYMQRYASKVSEGMALQLGCLELRRFFKDMPHNALDKKSNFELLEKEVGLDLFFPKQMQENLKPKQFRKMIQQTFQQYASLREEECVMKFFNTLAGFANIDQETYRCELIQGWNITVDLVIGPKGIRQLTSQDAKPTCLAEFKQIRSIRCLPLEEGQAVLQLGIEGAPQALSIKTSSLAEAENMADLIDGYCRLQGEHQGSLIIHPRKDGEKRNSLPQIPMLNLEARRSHLSESCSIESDIYAEIPDETLRRPGGPQYGIAREDVVLNRILGEGFFGEVYEGVYTNHKGEKINVAVKTCKKDCTLDNKEKFMSEAVIMKNLDHPHIVKLIGIIEEEPTWIIMELYPYGELGHYLERNKNSLKVLTLVLYSLQICKAMAYLESINCVHRDIAVRNILVASPECVKLGDFGLSRYIEDEDYYKASVTRLPIKWMSPESINFRRFTTASDVWMFAVCMWEILSFGKQPFFWLENKDVIGVLEKGDRLPKPDLCPPVLYTLMTRCWDYDPSDRPRFTELVCSLSDVYQMEKDIAMEQERNARYRTPKILEPTAFQEPPPKPSRPKYRPPPQTNLLAPKLQFQVPEGLCASSPTLTSPMEYPSPVNSLHTPPLHRHNVFKRHSMREEDFIQPSSREEAQQLWEAEKVKMRQILDKQQKQMVEDYQWLRQEEKSLDPMVYMNDKSPLTPEKEVGYLEFTGPPQKPPRLGAQSIQPTANLDRTDDLVYLNVMELVRAVLELKNELCQLPPEGYVVVVKNVGLTLRKLIGSVDDLLPSLPSSSRTEIEGTQKLLNKDLAELINKMRLAQQNAVTSLSEECKRQMLTASHTLAVDAKNLLDAVDQAKVLANLAHPPAE fak1_human Focal adhesion kinase 1 PTK2 FAK FAK1 pp125FAK Protein-tyrosine kinase 2 Focal adhesion kinase-related nonkinase Protein phosphatase 1 regulatory subunit 71 p125FAK protein protein human Homo sapiens Human MAAAYLDPNLNHTPNSSTKTHLGTGMERSPGAMERVLKVFHYFESNSEPTTWASIIRHGDATDVRGIIQKIVDSHKVKHVACYGFRLSHLRSEEVHWLHVDMGVSSVREKYELAHPPEEWKYELRIRYLPKGFLNQFTEDKPTLNFFYQQVKSDYMLEIADQVDQEIALKLGCLEIRRSYWEMRGNALEKKSNYEVLEKDVGLKRFFPKSLLDSVKAKTLRKLIQQTFRQFANLNREESILKFFEILSPVYRFDKECFKCALGSSWIISVELAIGPEEGISYLTDKGCNPTHLADFTQVQTIQYSNSEDKDRKGMLQLKIAGAPEPLTVTAPSLTIAENMADLIDGYCRLVNGTSQSFIIRPQKEGERALPSIPKLANSEKQGMRTHAVSVSETDDYAEIIDEEDTYTMPSTRDYEIQRERIELGRCIGEGQFGDVHQGIYMSPENPALAVAIKTCKNCTSDSVREKFLQEALTMRQFDHPHIVKLIGVITENPVWIIMELCTLGELRSFLQVRKYSLDLASLILYAYQLSTALAYLESKRFVHRDIAARNVLVSSNDCVKLGDFGLSRYMEDSTYYKASKGKLPIKWMAPESINFRRFTSASDVWMFGVCMWEILMHGVKPFQGVKNNDVIGRIENGERLPMPPNCPPTLYSLMTKCWAYDPSRRPRFTELKAQLSTILEEEKAQQEERMRMESRRQATVSWDSGGSDEAPPKPSRPGYPSPRSSEGFYPSPQHMVQTNHYQVSGYPGSHGITAMAGSIYPGQASLLDQTDSWNHRPQEIAMWQPNVEDSTVLDLRGIGQVLPTHLMEERLIRQQQEMEEDQRWLEKEERFLKPDVRLSRGSIDREDGSLQGPIGNQHIYQPVGKPDPAAPPKKPPRPGAPGHLGSLASLSSPADSYNEGVKLQPQEISPPPTANLDRSNDKVYENVTGLVKAVIEMSSKIQPAPPEEYVPMVKEVGLALRTLLATVDETIPLLPASTHREIEMAQKLLNSDLGELINKMKLAQQYVMTSLQQEYKKQMLTAAHALAVDAKNLLDVIDQARLKMLGQTRPH pak1_human Serine/threonine-protein kinase PAK 1 PAK1 p21-activated kinase 1 p65-PAK Alpha-PAK protein protein human Homo sapiens Human MSNNGLDIQDKPPAPPMRNTSTMIGAGSKDAGTLNHGSKPLPPNPEEKKKKDRFYRSILPGDKTNKKKEKERPEISLPSDFEHTIHVGFDAVTGEFTGMPEQWARLLQTSNITKSEQKKNPQAVLDVLEFYNSKKTSNSQKYMSFTDKSAEDYNSSNALNVKAVSETPAVPPVSEDEDDDDDDATPPPVIAPRPEHTKSVYTRSVIEPLPVTPTRDVATSPISPTENNTTPPDALTRNTEKQKKKPKMSDEEILEKLRSIVSVGDPKKKYTRFEKIGQGASGTVYTAMDVATGQEVAIKQMNLQQQPKKELIINEILVMRENKNPNIVNYLDSYLVGDELWVVMEYLAGGSLTDVVTETCMDEGQIAAVCRECLQALEFLHSNQVIHRDIKSDNILLGMDGSVKLTDFGFCAQITPEQSKRSTMVGTPYWMAPEVVTRKAYGPKVDIWSLGIMAIEMIEGEPPYLNENPLRALYLIATNGTPELQNPEKLSAIFRDFLNRCLEMDVEKRGSAKELLQHQFLKIAKPLSSLTPLIAAAKEATKNNH nck2_human Cytoplasmic protein NCK2 NCK adaptor protein 2 SH2/SH3 adaptor protein NCK-beta NCK2 GRB4 Growth factor receptor-bound protein 4 protein protein human Homo sapiens Human MTEEVIVIAKWDYTAQQDQELDIKKNERLWLLDDSKTWWRVRNAANRTGYVPSNYVERKNSLKKGSLVKNLKDTLGLGKTRRKTSARDASPTPSTDAEYPANGSGADRIYDLNIPAFVKFAYVAEREDELSLVKGSRVTVMEKCSDGWWRGSYNGQIGWFPSNYVLEEVDEAAAESPSFLSLRKGASLSNGQGSRVLHVVQTLYPFSSVTEEELNFEKGETMEVIEKPENDPEWWKCKNARGQVGLVPKNYVVVLSDGPALHPAHAPQISYTGPSSSGRFAGREWYYGNVTRHQAECALNERGVEGDFLIRDSESSPSDFSVSLKASGKNKHFKVQLVDNVYCIGQRRFHTMDELVEHYKKAPIFTSEHGEKLYLVRALQ rhou-ptk2b-2 RHOU (WRCH1) coimmunoprecipitates PTK2B (PYK2) in HEK-293T lysates 756692 N/A 756697 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 false Q7L0Q8:p.Thr63Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false n-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false Q7L0Q8:p.Pro80Gly mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q7L0Q8:p.Phe83Gly mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q7L0Q8:p.Phe86Cys mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false N/A 756698 unspecified role unspecified role prey prey physical association physical association false false false 2 C, 3 A, 4 A, 7 A, B PTK2B (PYK2) bound to the constitutively active mutant of RHOU (Q107L). Q14289; Q7L0Q8 transfection The mutations, P80G, F83G and F86C, are in the effector loop of RHOU (WRCH1). The immunoprecipitated RHOU (WRCH1) and PTK2B (PYK2) were tyrosine-phosphorylated. Kinase dead PTK2B (PYK2) bound to RHOU (WRCH1) indicating that kinase activity is not needed for this interaction. Treatment of cells with the Src inhibitor SU6656 blocked this interaction. Note this interacton did not occur in MEFs lacking Src,Yes and FYn. rhou-ptk2 RHOU (WRCH1) coimmunoprecipitates PTK2 (FAK) in HEK-293T lysates 756692 N/A 756697 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 false n-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false Q7L0Q8:p.Thr63Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q7L0Q8:p.Gln107Leu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false N/A 756699 unspecified role unspecified role prey prey physical association physical association false false false 2 D, 4 B Q05397; Q7L0Q8 transfection This interaction was much weaker than that between RHOU (WRCH1) and PTK2B (PYK2). PTK2 (FAK) interacted with the T63N mutant unlike PTK2B. The immunoprecipitated FAK was tyrosine-phosphorylated but RHOU (WRCH1) was not. The Q107LDeltaN Wrch1 construct interaction with FAK was weaker than the interaction of the Q107L construct with FAK. rhou-pak1 RHOU (WRCH1) coimmunoprecipitates PAK1 in HEK-293T lysates 756693 N/A 756697 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 false Q7L0Q8:p.Pro80Gly mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false Q7L0Q8:p.Phe86Cys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false N/A 756700 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 3 A Q13153; Q7L0Q8 transfection The mutations, P80G, F83G and F86C, are in the effector loop of RHOU (WRCH1). rhou-nck2 RHOU (WRCH1) coimmunoprecipitates NCK2 in HEK-293T lysates 756693 N/A 756697 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 false N/A 756701 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 3 A O43639; Q7L0Q8 transfection The authors comment that Nck2, which requires the Wrch1 N-terminal domain for binding, bound with equal affinity to all three effector domain mutants. rhou-ptk2b RHOU (WRCH1) interacts with PTK2B (PYK2) 756694 N/A 756698 unspecified role unspecified role prey prey N/A 756697 unspecified role unspecified role bait bait physical association physical association false false false The authors state the clones encoding PTK2B (PYK2) were of different lengths but all started in the N-terminal part of the FERM domain (given as aa 39-359 in the UniProtKB entry). However, the authors do not provide the actual ranges. rhou-ptk2b-1 RHOU (WRCH1) interacts with PTK2B (PYK2) 756695 N/A 756698 unspecified role unspecified role prey prey N/A 756697 unspecified role unspecified role bait bait Q7L0Q8:p.Thr63Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false n-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false 2 A The RHOU (WRCH1) construct with the proline-rich region (aa 1-46) deleted did not interact with PTK2B (PYK2). The authors deduce that RHOU (WRCH1) must be in the GTP-bound form as the putatively GDP-bound mutant (T63N) did not interact with PTK2B (PYK2). ptk2b-rhou RHOU (WRCH1) interacts with PTK2B (PYK2) in a pull down assay 756696 N/A 756697 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 false 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. N/A 756698 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 false region FERM sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range false physical association physical association false false false 2 E The coordinates for the FERM domain of PTK2B (PYK2) have been taken from the UniProtKB entry for the protein, as not provided by the authors, and thus fuzzy coordinates have been used.