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/ Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca 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 JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca Accepted 2008-FEB-31 by ORCHARD. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca ecoli Escherichia coli (strain K12) fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca Accepted 2008-FEB-31 by ORCHARD. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 ORCHARD Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca Accepted 2008-FEB-31 by ORCHARD. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca Accepted 2008-FEB-31 by ORCHARD. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 bifc bimolecular fluorescence complementation bifc tag fluorescence tag visualisation by fluorescence tag fluorescence JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca Accepted 2008-FEB-31 by ORCHARD. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 ORCHARD Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Dual expression recombinase based (DERB) single vector system for high throughput screening and verification of protein interaction in living cells. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca ecoli Escherichia coli (strain K12) fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence JianPing Lu, Jehonathan H Pinthus 2008-01-29: Lu JP, Juravinski Cancer Centre, Hamilton, Ontario, Canada. Protein interactions detected in the DERB platform with BiFC were validated by flow cytometery Jehonathan.Pinthus@hrcc.on.ca,jianping.lu@hrcc.on.ca Accepted 2008-FEB-31 by ORCHARD. Nature Precedings (http://precedings.nature.com/documents/1550/version/1) 2008 vime_human Vimentin VIM protein protein human Homo sapiens Human MSTRSVSSSSYRRMFGGPGTASRPSSSRSYVTTSTRTYSLGSALRPSTSRSLYASSPGGVYATRSSAVRLRSSVPGVRLLQDSVDFSLADAINTEFKNTRTNEKVELQELNDRFANYIDKVRFLEQQNKILLAELEQLKGQGKSRLGDLYEEEMRELRRQVDQLTNDKARVEVERDNLAEDIMRLREKLQEEMLQREEAENTLQSFRQDVDNASLARLDLERKVESLQEEIAFLKKLHEEEIQELQAQIQEQHVQIDVDVSKPDLTAALRDVRQQYESVAAKNLQEAEEWYKSKFADLSEAANRNNDALRQAKQESTEYRRQVQSLTCEVDALKGTNESLERQMREMEENFAVEAANYQDTIGRLQDEIQNMKEEMARHLREYQDLLNVKMALDIEIATYRKLLEGEESRISLPLPNFSSLNLRETNLDSLPLVDTHSKRTLLIKTVETRDGQVINETSQHHDDLE DIP protein P08670 original sequence version: 195 DIP protein P08670 original sequence version: 209 DIP protein P08670 original sequence version: 192 DIP protein P08670 original sequence version: 175 BAB54026665B015A paqr1_human Adiponectin receptor protein 1 ADIPOR1 PAQR1 CGI-45 Progestin and adipoQ receptor family member I TESBP1A Progestin and adipoQ receptor family member 1 protein protein human Homo sapiens Human MSSHKGSVVAQGNGAPASNREADTVELAELGPLLEEKGKRVIANPPKAEEEQTCPVPQEEEEEVRVLTLPLQAHHAMEKMEEFVYKVWEGRWRVIPYDVLPDWLKDNDYLLHGHRPPMPSFRACFKSIFRIHTETGNIWTHLLGFVLFLFLGILTMLRPNMYFMAPLQEKVVFGMFFLGAVLCLSFSWLFHTVYCHSEKVSRTFSKLDYSGIALLIMGSFVPWLYYSFYCSPQPRLIYLSIVCVLGISAIIVAQWDRFATPKHRQTRAGVFLGLGLSGVVPTMHFTIAEGFVKATTVGQMGWFFLMAVMYITGAGLYAARIPERFFPGKFDIWFQSHQIFHVLVVAAAFVHFYGVSNLQEFRYGLEGGCTDDTLL DIP protein Q96A54 original sequence version: 66 1CC0300A7D178EB0 dp13a_human DCC-interacting protein 13-alpha Adapter protein containing PH domain, PTB domain and leucine zipper motif 1 APPL1 APPL DIP13A KIAA1428 protein protein human Homo sapiens Human MPGIDKLPIEETLEDSPQTRSLLGVFEEDATAISNYMNQLYQAMHRIYDAQNELSAATHLTSKLLKEYEKQRFPLGGDDEVMSSTLQQFSKVIDELSSCHAVLSTQLADAMMFPITQFKERDLKEILTLKEVFQIASNDHDAAINRYSRLSKKRENDKVKYEVTEDVYTSRKKQHQTMMHYFCALNTLQYKKKIALLEPLLGYMQAQISFFKMGSENLNEQLEEFLANIGTSVQNVRREMDSDIETMQQTIEDLEVASDPLYVPDPDPTKFPVNRNLTRKAGYLNARNKTGLVSSTWDRQFYFTQGGNLMSQARGDVAGGLAMDIDNCSVMAVDCEDRRYCFQITSFDGKKSSILQAESKKDHEEWICTINNISKQIYLSENPEETAARVNQSALEAVTPSPSFQQRHESLRPAAGQSRPPTARTSSSGSLGSESTNLAALSLDSLVAPDTPIQFDIISPVCEDQPGQAKAFGQGGRRTNPFGESGGSTKSETEDSILHQLFIVRFLGSMEVKSDDHPDVVYETMRQILAARAIHNIFRMTESHLLVTCDCLKLIDPQTQVTRLTFPLPCVVLYATHQENKRLFGFVLRTSSGRSESNLSSVCYIFESNNEGEKICDSVGLAKQIALHAELDRRASEKQKEIERVKEKQQKELNKQKQIEKDLEEQSRLIAASSRPNQASSEGQFVVLSSSQSEESDLGEGGKKRESEA DIP protein Q9UKG1 original sequence version: 44 4CABECFCF4BB110D paqr2_human Adiponectin receptor protein 2 ADIPOR2 PAQR2 Progestin and adipoQ receptor family member 2 Progestin and adipoQ receptor family member II protein protein human Homo sapiens Human MNEPTENRLGCSRTPEPDIRLRKGHQLDGTRRGDNDSHQGDLEPILEASVLSSHHKKSSEEHEYSDEAPQEDEGFMGMSPLLQAHHAMEKMEEFVCKVWEGRWRVIPHDVLPDWLKDNDFLLHGHRPPMPSFRACFKSIFRIHTETGNIWTHLLGCVFFLCLGIFYMFRPNISFVAPLQEKVVFGLFFLGAILCLSFSWLFHTVYCHSEGVSRLFSKLDYSGIALLIMGSFVPWLYYSFYCNPQPCFIYLIVICVLGIAAIIVSQWDMFATPQYRGVRAGVFLGLGLSGIIPTLHYVISEGFLKAATIGQIGWLMLMASLYITGAALYAARIPERFFPGKCDIWFHSHQLFHIFVVAGAFVHFHGVSNLQEFRFMIGGGCSEEDAL B3B0FF0BBF118AA1 adipor1-adipor1 3 8 unspecified role unspecified role neutral component neutral component n terminal yfp 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 8 unspecified role unspecified role neutral component neutral component c terminal yfp 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 physical association physical association Investigation of pFER-YcADIPOR1-YnADIPOR1 with a 96-well plate reader and flow cytometer confirmed results identified with FRET procaryotic construct (Fig.5b) 5B adipor1-adipor1-1 6 8 unspecified role unspecified role neutral component neutral component region 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 8 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction 5A FRET investigation with pT7-ECFPADIPOR1-EYFPADIPOR1 resulted in the identification of a novel receptor interaction adipor1-appl1-1 1 8 unspecified role unspecified role bait bait region 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 9 unspecified role unspecified role prey prey region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association APPL1 presence within the ADIPOR1 collection and confirmed interaction 4 adipor2-appl1 6 10 unspecified role unspecified role neutral component neutral component region 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 9 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction 2a Constructs p-ECFPAPPL1-EYFPADIPOR1 and p-ECFPAPPL1-EYFPADIPOR2 displayed positive interaction FRETN mirroring the fusion value adipor1-appl1-3 4 8 unspecified role unspecified role neutral component neutral component region 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 9 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction Flow cytometry further validated positive interactions by detecting EYFP fluorescent cell populations in pFer-YcAPPL1-YnADIPOR1 transfected HeLa cells 2B adipor1-appl1-2 6 8 unspecified role unspecified role neutral component neutral component region 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 9 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction 2a Constructs p-ECFPAPPL1-EYFPADIPOR1 and p-ECFPAPPL1-EYFPADIPOR2 displayed positive interaction FRETN mirroring the fusion value adipor2-appl1-1 4 9 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role neutral component neutral component region 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 proximity proximity close-contact colocalization neighbourhood interaction 2 Constructs p-ECFPAPPL1-EYFPADIPOR1 and p-ECFPAPPL1-EYFPADIPOR2 displayed positive interaction FRETN mirroring the fusion value vim-vim-2 6 7 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role neutral component neutral component region 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 proximity proximity close-contact colocalization neighbourhood interaction 5A vim-vim-3 3 7 unspecified role unspecified role neutral component neutral component n terminal yfp 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 7 unspecified role unspecified role neutral component neutral component c terminal yfp 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 physical association physical association 3 Expression vector pFer-YcVIM-YnVIM transfected cells generated substantial EYFP emission above the negligible pFer-YcVIM-YnTUBA1B background in all analyzers. adipor1-appl1-4 3 8 unspecified role unspecified role neutral component neutral component n terminal yfp 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 9 unspecified role unspecified role neutral component neutral component c terminal yfp 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 physical association physical association 3 Flow cytometry further validated positive interactions by detecting EYFP fluorescent cell populations in pFer-YcAPPL1-YnADIPOR1 transfected HeLa cells adipor1-appl1-5 2 8 unspecified role unspecified role neutral component neutral component region 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 9 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction 2b Constructs p-ECFPAPPL1-EYFPADIPOR1 and p-ECFPAPPL1-EYFPADIPOR2 displayed positive interaction FRETN mirroring the fusion value adipor2-appl1-2 2 10 unspecified role unspecified role neutral component neutral component region 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 9 unspecified role unspecified role neutral component neutral component region cfp tag cyan fluorescent protein tag CFP cyan fluorescent protein cyan fluorescent protein CFP cfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined proximity proximity close-contact colocalization neighbourhood interaction 2b Constructs p-ECFPAPPL1-EYFPADIPOR1 and p-ECFPAPPL1-EYFPADIPOR2 displayed positive interaction FRETN mirroring the fusion value adipor1-adipor1-2 5 8 unspecified role unspecified role neutral component neutral component n terminal yfp 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 8 unspecified role unspecified role neutral component neutral component c terminal yfp 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 physical association physical association affirmation of the protein interaction through fluorescent microscopy with the pFer-YcADIPOR1-YnADIPOR1 expression vector 5B vim-vim-4 5 7 unspecified role unspecified role neutral component neutral component n terminal yfp 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 7 unspecified role unspecified role neutral component neutral component c terminal yfp 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 physical association physical association 3B Consecutive confirmation with fluroscent microscopy demonstrated interactions among pFer-YcVIM-ynVIM, pFer-YcAPPL1-YnADIPOR1 protein partners (Fig.3B) adipor1-appl1-6 5 8 unspecified role unspecified role neutral component neutral component n terminal yfp 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 9 unspecified role unspecified role neutral component neutral component c terminal yfp 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 physical association physical association 3B Consecutive confirmation with fluroscent microscopy demonstrated interactions among pFer-YcVIM-ynVIM, pFer-YcAPPL1-YnADIPOR1 protein partners (Fig.3B)