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/ Human glutathione S-transferase P1-1 interacts with TRAF2 and regulates TRAF2-ASK1 signals. Human glutathione S-transferase P1-1 interacts with TRAF2 and regulates TRAF2-ASK1 signals. Oncogene (0950-9232) 2006 imex curation Wu Y., Fan Y., Xue B., Luo L., Shen J., Zhang S., Jiang Y., Yin Z. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology Cells were untreated or treated with TNF alpha (50ng per ml) prior to assay. yinzhimin@njnu.edu.cn 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Wu Y., Fan Y., Xue B., Luo L., Shen J., Zhang S., Jiang Y., Yin Z. Apoptosis - Interactions involving proteins with a function related to apoptosis Cells were untreated or treated with TNF alpha (50ng per ml) prior to assay. yinzhimin@njnu.edu.cn Accepted 2006-SEP-15 by ORCHARD. 24-JAN-2007: Contacted by JYOTI. Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation Human glutathione S-transferase P1-1 interacts with TRAF2 and regulates TRAF2-ASK1 signals. Human glutathione S-transferase P1-1 interacts with TRAF2 and regulates TRAF2-ASK1 signals. Oncogene (0950-9232) 2006 imex curation Wu Y., Fan Y., Xue B., Luo L., Shen J., Zhang S., Jiang Y., Yin Z. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology Cells were untreated or treated with TNF alpha (50ng per ml) prior to assay. yinzhimin@njnu.edu.cn 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Wu Y., Fan Y., Xue B., Luo L., Shen J., Zhang S., Jiang Y., Yin Z. Apoptosis - Interactions involving proteins with a function related to apoptosis Cells were untreated or treated with TNF alpha (50ng per ml) prior to assay. yinzhimin@njnu.edu.cn Accepted 2006-SEP-15 by ORCHARD. Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation Human glutathione S-transferase P1-1 interacts with TRAF2 and regulates TRAF2-ASK1 signals. Human glutathione S-transferase P1-1 interacts with TRAF2 and regulates TRAF2-ASK1 signals. Oncogene (0950-9232) 2006 imex curation Wu Y., Fan Y., Xue B., Luo L., Shen J., Zhang S., Jiang Y., Yin Z. Apoptosis - Interactions involving proteins with a function related to apoptosis BioCreative - Critical Assessment of Information Extraction systems in Biology Cells were untreated or treated with TNF alpha (50ng per ml) prior to assay. yinzhimin@njnu.edu.cn 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Wu Y., Fan Y., Xue B., Luo L., Shen J., Zhang S., Jiang Y., Yin Z. Apoptosis - Interactions involving proteins with a function related to apoptosis Cells were untreated or treated with TNF alpha (50ng per ml) prior to assay. yinzhimin@njnu.edu.cn Accepted 2006-SEP-15 by ORCHARD. 24-JAN-2007: Contacted by JYOTI. Oncogene (0950-9232) 2006 Only protein-protein interactions imex curation gstp1_human Glutathione S-transferase P GSTP1 GST3 FAEES3 GST class-pi GSTP1-1 protein protein human Homo sapiens Human MPPYTVVYFPVRGRCAALRMLLADQGQSWKEEVVTVETWQEGSLKASCLYGQLPKFQDGDLTLYQSNTILRHLGRTLGLYGKDQQEAALVDMVNDGVEDLRCKYISLIYTNYEAGKDDYVKALPGQLKPFETLLSQNQGGKTFIVGDQISFADYNLLDLLLIHEVLAPGCLDAFPLLSAYVGRLSARPKLKAFLASPEYVNLPINGNGKQ 409E33FFAA338396 m3k1_human Mitogen-activated protein kinase kinase kinase 1 MAPK/ERK kinase kinase 1 MAP3K1 MAPKKK1 MEKK MEKK1 protein protein human Homo sapiens Human MAAAAGNRASSSGFPGARATSPEAGGGGGALKASSAPAAAAGLLREAGSGGRERADWRRRQLRKVRSVELDQLPEQPLFLAASPPASSTSPSPEPADAAGSGTGFQPVAVPPPHGAASRGGAHLTESVAAPDSGASSPAAAEPGEKRAPAAEPSPAAAPAGREMENKETLKGLHKMDDRPEERMIREKLKATCMPAWKHEWLERRNRRGPVVVKPIPVKGDGSEMNHLAAESPGEVQASAASPASKGRRSPSPGNSPSGRTVKSESPGVRRKRVSPVPFQSGRITPPRRAPSPDGFSPYSPEETNRRVNKVMRARLYLLQQIGPNSFLIGGDSPDNKYRVFIGPQNCSCARGTFCIHLLFVMLRVFQLEPSDPMLWRKTLKNFEVESLFQKYHSRRSSRIKAPSRNTIQKFVSRMSNSHTLSSSSTSTSSSENSIKDEEEQMCPICLLGMLDEESLTVCEDGCRNKLHHHCMSIWAEECRRNREPLICPLCRSKWRSHDFYSHELSSPVDSPSSLRAAQQQTVQQQPLAGSRRNQESNFNLTHYGTQQIPPAYKDLAEPWIQVFGMELVGCLFSRNWNVREMALRRLSHDVSGALLLANGESTGNSGGSSGSSPSGGATSGSSQTSISGDVVEACCSVLSMVCADPVYKVYVAALKTLRAMLVYTPCHSLAERIKLQRLLQPVVDTILVKCADANSRTSQLSISTLLELCKGQAGELAVGREILKAGSIGIGGVDYVLNCILGNQTESNNWQELLGRLCLIDRLLLEFPAEFYPHIVSTDVSQAEPVEIRYKKLLSLLTFALQSIDNSHSMVGKLSRRIYLSSARMVTTVPHVFSKLLEMLSVSSSTHFTRMRRRLMAIADEVEIAEAIQLGVEDTLDGQQDSFLQASVPNNYLETTENSSPECTVHLEKTGKGLCATKLSASSEDISERLASISVGPSSSTTTTTTTTEQPKPMVQTKGRPHSQCLNSSPLSHHSQLMFPALSTPSSSTPSVPAGTATDVSKHRLQGFIPCRIPSASPQTQRKFSLQFHRNCPENKDSDKLSPVFTQSRPLPSSNIHRPKPSRPTPGNTSKQGDPSKNSMTLDLNSSSKCDDSFGCSSNSSNAVIPSDETVFTPVEEKCRLDVNTELNSSIEDLLEASMPSSDTTVTFKSEVAVLSPEKAENDDTYKDDVNHNQKCKEKMEAEEEEALAIAMAMSASQDALPIVPQLQVENGEDIIIIQQDTPETLPGHTKAKQPYREDTEWLKGQQIGLGAFSSCYQAQDVGTGTLMAVKQVTYVRNTSSEQEEVVEALREEIRMMSHLNHPNIIRMLGATCEKSNYNLFIEWMAGGSVAHLLSKYGAFKESVVINYTEQLLRGLSYLHENQIIHRDVKGANLLIDSTGQRLRIADFGAAARLASKGTGAGEFQGQLLGTIAFMAPEVLRGQQYGRSCDVWSVGCAIIEMACAKPPWNAEKHSNHLALIFKIASATTAPSIPSHLSPGLRDVALRCLELQPQDRPPSRELLKHPVFRTTW 5CB78242295411D9 m3k5_human Mitogen-activated protein kinase kinase kinase 5 MAPK/ERK kinase kinase 5 Apoptosis signal-regulating kinase 1 MAP3K5 ASK1 MAPKKK5 MEKK5 protein protein human Homo sapiens Human MSTEADEGITFSVPPFAPSGFCTIPEGGICRRGGAAAVGEGEEHQLPPPPPGSFWNVESAAAPGIGCPAATSSSSATRGRGSSVGGGSRRTTVAYVINEASQGQLVVAESEALQSLREACETVGATLETLHFGKLDFGETTVLDRFYNADIAVVEMSDAFRQPSLFYHLGVRESFSMANNIILYCDTNSDSLQSLKEIICQKNTMCTGNYTFVPYMITPHNKVYCCDSSFMKGLTELMQPNFELLLGPICLPLVDRFIQLLKVAQASSSQYFRESILNDIRKARNLYTGKELAAELARIRQRVDNIEVLTADIVINLLLSYRDIQDYDSIVKLVETLEKLPTFDLASHHHVKFHYAFALNRRNLPGDRAKALDIMIPMVQSEGQVASDMYCLVGRIYKDMFLDSNFTDTESRDHGASWFKKAFESEPTLQSGINYAVLLLAAGHQFESSFELRKVGVKLSSLLGKKGNLEKLQSYWEVGFFLGASVLANDHMRVIQASEKLFKLKTPAWYLKSIVETILIYKHFVKLTTEQPVAKQELVDFWMDFLVEATKTDVTVVRFPVLILEPTKIYQPSYLSINNEVEEKTISIWHVLPDDKKGIHEWNFSASSVRGVSISKFEERCCFLYVLHNSDDFQIYFCTELHCKKFFEMVNTITEEKGRSTEEGDCESDLLEYDYEYDENGDRVVLGKGTYGIVYAGRDLSNQVRIAIKEIPERDSRYSQPLHEEIALHKHLKHKNIVQYLGSFSENGFIKIFMEQVPGGSLSALLRSKWGPLKDNEQTIGFYTKQILEGLKYLHDNQIVHRDIKGDNVLINTYSGVLKISDFGTSKRLAGINPCTETFTGTLQYMAPEIIDKGPRGYGKAADIWSLGCTIIEMATGKPPFYELGEPQAAMFKVGMFKVHPEIPESMSAEAKAFILKCFEPDPDKRACANDLLVDEFLKVSSKKKKTQPKLSALSAGSNEYLRSISLPVPVLVEDTSSSSEYGSVSPDTELKVDPFSFKTRAKSCGERDVKGIRTLFLGIPDENFEDHSAPPSPEEKDSGFFMLRKDSERRATLHRILTEDQDKIVRNLMESLAQGAEEPKLKWEHITTLIASLREFVRSTDRKIIATTLSKLKLELDFDSHGISQVQVVLFGFQDAVNKVLRNHNIKPHWMFALDSIIRKAVQTAITILVPELRPHFSLASESDTADQEDLDVEDDHEEQPSNQTVRRPQAVIEDAVATSGVSTLSSTVSHDSQSAHRSLNVQLGRMKIETNRLLEELVRKEKELQALLHRAIEEKDQEIKHLKLKSQPIEIPELPVFHLNSSGTNTEDSELTDWLRVNGADEDTISRFLAEDYTLLDVLYYVTRDDLKCLRLRGGMLCTLWKAIIDFRNKQT DIP protein Q99683 original sequence version: 83 DIP protein Q99683 original sequence version: 151 DIP protein Q99683 original sequence version: 113 265BDC65968AF985 mk14_human Mitogen-activated protein kinase 14 Mitogen-activated protein kinase p38 alpha Cytokine suppressive anti-inflammatory drug-binding protein MAX-interacting protein 2 MAP kinase MXI2 SAPK2A MAPK14 CSBP CSBP1 CSBP2 CSPB1 MXI2 Stress-activated protein kinase 2a protein protein human Homo sapiens Human MSQERPTFYRQELNKTIWEVPERYQNLSPVGSGAYGSVCAAFDTKTGLRVAVKKLSRPFQSIIHAKRTYRELRLLKHMKHENVIGLLDVFTPARSLEEFNDVYLVTHLMGADLNNIVKCQKLTDDHVQFLIYQILRGLKYIHSADIIHRDLKPSNLAVNEDCELKILDFGLARHTDDEMTGYVATRWYRAPEIMLNWMHYNQTVDIWSVGCIMAELLTGRTLFPGTDHIDQLKLILRLVGTPGAELLKKISSESARNYIQSLTQMPKMNFANVFIGANPLAVDLLEKMLVLDSDKRITAAQALAHAYFAQYHDPDDEPVADPYDQSFESRDLLIDEWKSLTYDEVISFVPPPLDQEEMES 286C81D0487618B3 traf2_human TNF receptor-associated factor 2 TRAF2 TRAP3 Tumor necrosis factor type 2 receptor-associated protein 3 E3 ubiquitin-protein ligase TRAF2 RING-type E3 ubiquitin transferase TRAF2 protein protein human Homo sapiens Human MAAASVTPPGSLELLQPGFSKTLLGTKLEAKYLCSACRNVLRRPFQAQCGHRYCSFCLASILSSGPQNCAACVHEGIYEEGISILESSSAFPDNAARREVESLPAVCPSDGCTWKGTLKEYESCHEGRCPLMLTECPACKGLVRLGEKERHLEHECPERSLSCRHCRAPCCGADVKAHHEVCPKFPLTCDGCGKKKIPREKFQDHVKTCGKCRVPCRFHAIGCLETVEGEKQQEHEVQWLREHLAMLLSSVLEAKPLLGDQSHAGSELLQRCESLEKKTATFENIVCVLNREVERVAMTAEACSRQHRLDQDKIEALSSKVQQLERSIGLKDLAMADLEQKVLEMEASTYDGVFIWKISDFARKRQEAVAGRIPAIFSPAFYTSRYGYKMCLRIYLNGDGTGRGTHLSLFFVVMKGPNDALLRWPFNQKVTLMLLDQNNREHVIDAFRPDVTSSSFQRPVNDMNIASGCPLFCPVSKMEAKNSYVRDDAIFIKAIVDLTGL DIP protein Q12933 original sequence version: 131 DIP protein Q12933 original sequence version: 129 DIP protein Q12933 original sequence version: 167 DIP protein Q12933 original sequence version: 193 DIP protein Q12933-1 original sequence version: 154 DIP protein Q12933 original sequence version: 130 DIP protein Q12933 original sequence version: 151 C508BE185B783B20 gstp1-jnk1 2 4 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Figure 2a The results showed that GSTP1-1 associated with JNK1 in unstimulated cells, The results showed that GSTP1-1 associated with JNK1 in unstimulated cells, but binding decreased in response to Tumour Necrocis Factor alpha (TNF alpha) stimulus. TNF alpha gstp1-mapk13 2 4 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Figure 2a A weak interaction between GSTP1-1 and p38, which was unaffected by TNF, was also observed The results showed that GSTP1-1 associated with MAPK13 weakly in stimulated and untreated cells. gstp1-traf2 2 4 unspecified role unspecified role prey prey 8 unspecified role unspecified role bait bait physical association physical association Figure 2a Figure 3a The results from imunoblot analysis using anti-GSTP1-1 antibody of the TRAF2 of TRAF6 immunoprecipitates revealed that GSTP1-1 specifically associated with TRAF2 TNF alpha The amount of TRAF2 GSTP1-1 complex decreased to a minimum 45 minutes after treatment with TNF alpha (50nm per ml)and then returned to a basal level after 90 minutes. gstp1-traf2-2 1 4 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 8 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 Figure 2c Flag-TRAF2 but not Flag-TRAF6 specifically associated with GSTP1-1 gstp1-traf2-3 3 4 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 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 Figure 2d Moreover, in vitro pull-down assay using bacteria expressed recombinant His-GSTP1-1 and the HEK293 cell lysates containing Flag-tagged TRAF2 or TRAF6 showed the same result. gstp1-traf2-4 1 4 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 8 unspecified role unspecified role bait bait 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Figure 3b Mapping of TRAF2 domains required for GSTP1-1 binding traf2-ask1 1 6 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 8 unspecified role unspecified role bait bait 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 Figure 6 Figure 7 GSTP1 inhibits TRAF2-induced ASK1 catalytic activity. ASK1 was phosphorylated at threonine 845. GSTP1-1 (UniProt accession Q5TZY3). traf2-mekk1 1 8 unspecified role unspecified role bait bait 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 5 unspecified role unspecified role prey prey physical association physical association Figure 7 interaction between TRAF2 and MEKK1