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/ heckler-2013-1 Protein disulfide-isomerase interacts with soluble guanylyl cyclase via a redox-based mechanism and modulates its activity. rat-sv40lt-smc Rattus norvegicus sv40lt-smc SV40LT-SMC Rat aortic smooth muscle cells pla proximity ligation assay pELISA pELISA PLA antibody detection identification by antibody antibody detection Heckler E.J., Crassous P.A., Baskaran P., Beuve A. Affinomics - Interactions curated for the Affinomics consortium. Results from Fig 1 cannot be included as sGC (Bait) is a dimeric protein, - the beta subunit only being his tagged, but the PDI binding subunit is not known. (see reference 5) Similarly for Figure 2 (PDI catalysed thiol-disulfide exchange/reduction) which/both of the sGC subunits affected is not known. Cells used were ATCC (cat-no CRL-2018) rat aortic SMCs(smooth muscle cells); a sub-clone of SV40LT-SMC cells. The protein "PDI" was found to correspond to the gene P4hb/Pdia1 by mutation mapping. heckleer@njms.rutgers.edu heckleer@umdnj.edu Biochem. J. (0264-6021) 2013 Only protein-protein interactions imex curation heckler-2013-2 Protein disulfide-isomerase interacts with soluble guanylyl cyclase via a redox-based mechanism and modulates its activity. chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Heckler E.J., Crassous P.A., Baskaran P., Beuve A. Affinomics - Interactions curated for the Affinomics consortium. Results from Fig 1 cannot be included as sGC (Bait) is a dimeric protein, - the beta subunit only being his tagged, but the PDI binding subunit is not known. (see reference 5) Similarly for Figure 2 (PDI catalysed thiol-disulfide exchange/reduction) which/both of the sGC subunits affected is not known. COS-7 cells were transiently transfected with FLAG-tagged PDI WT or FLAG-tagged PDI CXXS and transfected with (rat) sGCα and sGCβ WT adenovirus. The protein "PDI" was found to correspond to the gene P4hb/Pdia1 by mutation mapping. heckleer@njms.rutgers.edu heckleer@umdnj.edu Biochem. J. (0264-6021) 2013 Only protein-protein interactions imex curation gcya1_rat Guanylate cyclase soluble subunit alpha-1 Gucy1a1 Guanylate cyclase soluble subunit alpha-3 Guc1a1 Gucy1a3 Soluble guanylate cyclase large subunit protein protein rat Rattus norvegicus (Rat) Rat MFCRKFKDLKITGECPFSLLAPGQVPTEPIEEVAGVSESCQATLPTCQEFAENAEGSHPQRKTSRNRVYLHTLAESIGKLIFPEFERLNLALQRTLAKHKIKENRNSSEKEDLERIIAEEAIAAGVPVEVLKDSLGEELFKICYEEDEHILGVVGGTLKDFLNSFSTLLKQSSHCQEAERRGRLEDASILCLDKDQDFLNVYYFFPKRTTALLLPGIIKAAARILYESHVEVSLMPPCFRSECTEFVNQPYLLYSVHVKSTKPSLSPGKPQSSLVIPTSLFCKTFPFHFMLDRDLAILQLGNGIRRLVNKRDFQGKPNFEEFFEILTPKINQTFSGIMTMLNMQFVIRVRRWDNLVKKSSRVMDLKGQMIYIVESSAILFLGSPCVDRLEDFTGRGLYLSDIPIHNALRDVVLIGEQARAQDGLKKRLGKLKATLEHAHQALEEEKKKTVDLLCSIFPSEVAQQLWQGQIVQAKKFNEVTMLFSDIVGFTAICSQCSPLQVITMLNALYTRFDQQCGELDVYKVETIGDAYCVAGGLHRESDTHAVQIALMALKMMELSNEVMSPHGEPIKMRIGLHSGSVFAGVVGVKMPRYCLFGNNVTLANKFESCSVPRKINVSPTTYRLLKDCPGFVFTPRSREELPPNFPSDIPGICHFLDAYQHQGPNSKPWFQQKDAEDGNANFLGKASGVD pdia1_rat Protein disulfide-isomerase Prolyl 4-hydroxylase subunit beta Cellular thyroid hormone-binding protein P4hb Pdia1 protein protein rat Rattus norvegicus (Rat) Rat MLSRALLCLALAWAARVGADALEEEDNVLVLKKSNFAEALAAHNYLLVEFYAPWCGHCKALAPEYAKAAAKLKAEGSEIRLAKVDATEESDLAQQYGVRGYPTIKFFKNGDTASPKEYTAGREADDIVNWLKKRTGPAATTLSDTAAAESLVDSSEVTVIGFFKDAGSDSAKQFLLAAEAVDDIPFGITSNSDVFSKYQLDKDGVVLFKKFDEGRNNFEGEITKEKLLDFIKHNQLPLVIEFTEQTAPKIFGGEIKTHILLFLPKSVSDYDGKLSNFKKAAEGFKGKILFIFIDSDHTDNQRILEFFGLKKEECPAVRLITLEEEMTKYKPESDELTAEKITQFCHHFLEGKIKPHLMSQELPEDWDKQPVKVLVGKNFEEVAFDEKKNVFVEFYAPWCGHCKQLAPIWDKLGETYKDHENIVIAKMDSTANEVEAVKVHSFPTLKFFPASADRTVIDYNGERTLDGFKKFLESGGQDGAGDNDDLDLEEALEPDMEEDDDQKAVKDEL gcyb1_rat Guanylate cyclase soluble subunit beta-1 Guanylate cyclase soluble subunit beta-3 Guc1b3 Gucy1b3 Gucy1b1 Soluble guanylate cyclase small subunit protein protein rat Rattus norvegicus (Rat) Rat MYGFVNHALELLVIRNYGPEVWEDIKKEAQLDEEGQFLVRIIYDDSKTYDLVAAASKVLNLNAGEILQMFGKMFFVFCQESGYDTILRVLGSNVREFLQNLDALHDHLATIYPGMRAPSFRCTDAEKGKGLILHYYSEREGLQDIVIGIIKTVAQQIHGTEIDMKVIQQRSEECDHTQFLIEEKESKEEDFYEDLDRFEENGTQDSRISPYTFCKAFPFHIIFDRDLVVTQCGNAIYRVLPQLQPGKCSLLSVFSLVRPHIDISFHGILSHINTVFVLRSKEGLLDVEKLECEDELTGAEISCLRLKGQMIYLPEADSILFLCSPSVMNLDDLTRRGLYLSDIPLHDATRDLVLLGEQFREEYKLTQELEILTDRLQLTLRALEDEKKKTDTLLYSVLPPSVANELRHKRPVPAKRYDNVTILFSGIVGFNAFCSKHASGEGAMKIVNLLNDLYTRFDTLTDSRKNPFVYKVETVGDKYMTVSGLPEPCIHHARSICHLALDMMEIAGQVQVDGESVQITIGIHTGEVVTGVIGQRMPRYCLFGNTVNLTSRTETTGEKGKINVSEYTYRCLMSPENSDPQFHLEHRGPVSMKGKKEPMQVWFLSRKNTGTEETNQDEN mint pdia1_mouse Protein disulfide-isomerase Prolyl 4-hydroxylase subunit beta Cellular thyroid hormone-binding protein p55 Endoplasmic reticulum resident protein 59 P4hb Pdia1 protein protein mouse Mus musculus Mouse MLSRALLCLALAWAARVGADALEEEDNVLVLKKSNFEEALAAHKYLLVEFYAPWCGHCKALAPEYAKAAAKLKAEGSEIRLAKVDATEESDLAQQYGVRGYPTIKFFKNGDTASPKEYTAGREADDIVNWLKKRTGPAATTLSDTAAAESLVDSSEVTVIGFFKDVESDSAKQFLLAAEAIDDIPFGITSNSGVFSKYQLDKDGVVLFKKFDEGRNNFEGEITKEKLLDFIKHNQLPLVIEFTEQTAPKIFGGEIKTHILLFLPKSVSDYDGKLSSFKRAAEGFKGKILFIFIDSDHTDNQRILEFFGLKKEECPAVRLITLEEEMTKYKPESDELTAEKITEFCHRFLEGKIKPHLMSQEVPEDWDKQPVKVLVGANFEEVAFDEKKNVFVEFYAPWCGHCKQLAPIWDKLGETYKDHENIIIAKMDSTANEVEAVKVHSFPTLKFFPASADRTVIDYNGERTLDGFKKFLESGGQDGAGDDEDLDLEEALEPDMEEDDDQKAVKDEL gcs_alpha_1-p4hb 1547399 n/a 1547401 unspecified role unspecified role neutral component neutral component n/a 1547402 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 5 gcs_alpha_1-p4hb-1 1547399 n/a 1547403 unspecified role unspecified role neutral component neutral component n/a 1547402 unspecified role unspecified role neutral component neutral component proximity proximity close-contact colocalization neighbourhood interaction false false false Figure 5i p4hb-gucy1b3 1547400 n/a 1547403 unspecified role unspecified role prey prey n/a 1547404 unspecified role unspecified role bait bait c_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Figure 3A FLAG-tagged PDI WT and PDI CXXS pulled down sGC in COS-7 lysates, as a complex of approximately 200 kDa and of higher molecular mass (potential multimers). Addition of reductant (2-mercaptoethanol) to the bound samples resolves the high-molecular-mass complexes to the expected sGCa and sGCb subunit size (~80 and 75 kDa), gcs_alpha_1-p4hb-2 1547400 n/a 1547404 unspecified role unspecified role bait bait c_terminus flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1547401 unspecified role unspecified role prey prey physical association physical association false false false Figure 3A FLAG-tagged PDI WT and PDI CXXS pulled down sGC in COS-7 lysates, as a complex of approximately 200 kDa and of higher molecular mass (potential multimers). Addition of reductant (2-mercaptoethanol) to the bound samples resolves the high-molecular-mass complexes to the expected sGCa and sGCb subunit size (~80 and 75 kDa),