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/ stephens-1997-1 Serine 331 and tyrosine 333 are both involved in the interaction between the cytosolic domain of TGN38 and the mu2 subunit of the AP2 clathrin adaptor complex. 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 Stephens DJ., Crump CM., Clarke AR., Banting G. J. Biol. Chem. (0021-9258) 1997 stephens-1997-2 Serine 331 and tyrosine 333 are both involved in the interaction between the cytosolic domain of TGN38 and the mu2 subunit of the AP2 clathrin adaptor complex. in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip predetermined predetermined participant predetermined Stephens DJ., Crump CM., Clarke AR., Banting G. interacting proteins were detected by measuring bands intensity after SDS-PAGE staining. J. Biol. Chem. (0021-9258) 1997 stephens-1997-3 Serine 331 and tyrosine 333 are both involved in the interaction between the cytosolic domain of TGN38 and the mu2 subunit of the AP2 clathrin adaptor complex. in vitro In vitro fluorescence spectr classical fluorescence spectroscopy fluorescence spectr predetermined predetermined participant predetermined Stephens DJ., Crump CM., Clarke AR., Banting G. J. Biol. Chem. (0021-9258) 1997 ap2m1_rat AP-2 complex subunit mu Adaptor protein complex AP-2 subunit mu AP-2 mu chain Ap2m1 Mu2-adaptin Plasma membrane adaptor AP-2 50 kDa protein Clathrin coat assembly protein AP50 Clathrin coat-associated protein AP50 Clathrin assembly protein complex 2 mu medium chain Adaptor-related protein complex 2 subunit mu protein protein rat Rattus norvegicus (Rat) Rat MIGGLFIYNHKGEVLISRVYRDDIGRNAVDAFRVNVIHARQQVRSPVTNIARTSFFHVKRSNIWLAAVTKQNVNAAMVFEFLYKMCDVMAAYFGKISEENIKNNFVLIYELLDEILDFGYPQNSETGALKTFITQQGIKSQHQTKEEQSQITSQVTGQIGWRREGIKYRRNELFLDVLESVNLLMSPQGQVLSAHVSGRVVMKSYLSGMPECKFGMNDKIVIEKQGKGTADETSKSGKQSIAIDDCTFHQCVRLSKFDSERSISFIPPDGEFELMRYRTTKDIILPFRVIPLVREVGRTKLEVKVVIKSNFKPSLLAQKIEVRIPTPLNTSGVQVICMKGKAKYKASENAIVWKIKRMAGMKESQISAEIELLPTNDKKKWARPPISMNFEVPFAPSGLKVRYLKVFEPKLNYSDHDVIKWVRYIGRSGIYETRC tgon3_rat Trans-Golgi network integral membrane protein TGN38 Ttgn1 protein protein rat Rattus norvegicus (Rat) Rat MQFLVALLLLSVAVARALPSASKPNNTSSENNPPIQPSTPLPPGVDISQQVKTNRPTDQRLESDKEGQDKTVARTSASVSSGVESATNLNLDDSKKHPETADAKLKETLQQLLPVDPKQEKSGQKFTKDSGSPTGGDSDNTTGGDSNKTTGVDSDKTSGGDSNKPTGSDNDKPTGGDSNKPTSKVPSNTETPKIDKVQLTEKGQKPTLISKTESGEKLAGDSDFSLKPEKGDKSSEPTEDVETKEIEEGDTEPEEGSPLEEENEKVLGPSSSENQEGTLTDSMKDEKDDHYKDNSGNTSAESSHFFAYLVTAAVLVAVLYIAYHNKRKIIAFALEGKRSKVTRRPKASDYQRLNLKL ap2m1-tgn41-3 Y2H: bait = TGN38 (cytoplasmic tail with the YQRL motif), prey = adaptin mu2 182723 N/A 182726 unspecified role unspecified role prey prey N/A 182727 unspecified role unspecified role bait bait P19814:p.Tyr350Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false cytoplasmic domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false P19814:p.Ser348Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false The tyrosine 350 and Serine 348 hotspotsare in the transport signal motif SDYQRL (348-353) ap2m1-tgn41-4 In vitro Co-IP: bait = TGN38 (cytoplasmic tail with the YQRL motif), prey = adaptin mu2 182724 N/A 182726 unspecified role unspecified role prey prey region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false thioredoxin tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) N/A 182727 unspecified role unspecified role bait bait P19814:p.Tyr350Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false cytoplasmic domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false P19814:p.Ser348Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false 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 ecobb Escherichia coli (strain B / BL21) physical association physical association false false false The tyrosine 350 and Serine 348 hotspots are in the transport signal motif SDYQRL (348-353) Both bait and prey are purified. ap2m1-tgn41-5 In vitro Co-IP: bait = TGN38 (cytoplasmic tail with the YQRL motif), prey = adaptin mu2 182725 N/A 182727 unspecified role unspecified role bait bait cytoplasmic domain binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false P19814:p.Ser348Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false 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 P19814:p.Tyr350Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) N/A 182726 unspecified role unspecified role prey prey region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false thioredoxin tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) physical association physical association false false false The tyrosine 350 and Serine 348 hotspots are in the transport signal motif SDYQRL (348-353) Both bait and prey are purified. The affinity between TGN38 cytoplasmic tail and the mutant Y350A adaptin mu2 mutant is kd = 7.55 10-7 M The affinity between TGN38 cytoplasmic tail and the mutant S348A adaptin mu2 mutant is kd = 1.03 10-7 M