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/ ren-2013-2 Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. james.hurley@nih.gov Cell (0092-8674) 2013 Only protein-protein interactions imex curation ren-2013-4 Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. canlf-mdck Canis familiaris (dog) kidney epithelial cells Canis lupus familiaris Dog Canis familiaris mdck Canine Cocker Spaniel kidney epithelial cell line anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. james.hurley@nih.gov Cell (0092-8674) 2013 Only protein-protein interactions imex curation ren-2013-5 Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 confocal microscopy confocal microscopy tag fluorescence tag visualisation by fluorescence tag fluorescence Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. Untagged proteins identified by immunostaining. james.hurley@nih.gov Cell (0092-8674) 2013 Only protein-protein interactions imex curation ren-2013-1 Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. james.hurley@nih.gov Cell (0092-8674) 2013 Only protein-protein interactions imex curation ren-2013-3 Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. in vitro In vitro fluorescence fluorescence technology fluorescence predetermined predetermined participant predetermined Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. Differential Scanning Fluorimetry (DSF) was used to measure the temperature at which the complex unfolded by an increase in the fluorescence of a dye (SYPRO orange) with affinity for hydrophobic parts of the proteins, which are exposed as the complex unfolds. james.hurley@nih.gov Cell (0092-8674) 2013 Only protein-protein interactions imex curation ren-2013-6 Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. james.hurley@nih.gov Cell (0092-8674) 2013 Only protein-protein interactions imex curation ap1m1_mouse AP-1 complex subunit mu-1 Ap1m1 Cltnm Golgi adaptor HA1/AP1 adaptin mu-1 subunit Mu-adaptin 1 AP-mu chain family member mu1A Mu1A-adaptin Clathrin coat assembly protein AP47 Clathrin coat-associated protein AP47 Adaptor protein complex AP-1 subunit mu-1 Clathrin assembly protein complex 1 mu-1 medium chain 1 Adaptor-related protein complex 1 subunit mu-1 protein protein mouse Mus musculus Mouse MSASAVYVLDLKGKVLICRNYRGDVDMSEVEHFMPILMEKEEEGMLSPILAHGGVRFMWIKHNNLYLVATSKKNACVSLVFSFLYKVVQVFSEYFKELEEESIRDNFVIIYELLDELMDFGYPQTTDSKILQEYITQEGHKLETGAPRPPATVTNAVSWRSEGIKYRKNEVFLDVIEAVNLLVSANGNVLRSEIVGSIKMRVFLSGMPELRLGLNDKVLFDNTGRGKSKSVELEDVKFHQCVRLSRFENDRTISFIPPDGEFELMSYRLNTHVKPLIWIESVIEKHSHSRIEYMVKAKSQFKRRSTANNVEIHIPVPNDADSPKFKTTVGSVKWVPENSEIVWSVKSFPGGKEYLMRAHFGLPSVEAEDKEGKPPISVKFEIPYFTTSGIQVRYLKIIEKSGYQALPWVRYITQNGDYQLRTQ ap1g1_mouse AP-1 complex subunit gamma-1 Ap1g1 Adtg Clapg1 Adaptor protein complex AP-1 subunit gamma-1 Golgi adaptor HA1/AP1 adaptin subunit gamma-1 Gamma1-adaptin Gamma-adaptin Clathrin assembly protein complex 1 gamma-1 large chain Adaptor-related protein complex 1 subunit gamma-1 protein protein mouse Mus musculus Mouse MPAPIRLRELIRTIRTARTQAEEREMIQKECAAIRSSFREEDNTYRCRNVAKLLYMHMLGYPAHFGQLECLKLIASQKFTDKRIGYLGAMLLLDERQDVHLLMTNCIKNDLNHSTQFVQGLALCTLGCMGSSEMCRDLAGEVEKLLKTSNSYLRKKAALCAVHVIRKVPELMEMFLPATKNLLNEKNHGVLHTSVVLLTEMCERSPDMLAHFRKLVPQLVRILKNLIMSGYSPEHDVSGISDPFLQVRILRLLRILGRNDDDSSEAMNDILAQVATNTETSKNVGNAILYETVLTIMDIKSESGLRVLAINILGRFLLNNDKNIRYVALTSLLKTVQTDHNAVQRHRSTIVDCLKDLDVSIKRRAMELSFALVNGNNIRGMMKELLYFLDSCEPEFKADCASGIFLAAEKYAPSKRWHIDTIMRVLTTAGSYVRDDAVPNLIQLITNSVEMHAYTVQRLYKAILGDYSQQPLVQVAAWCIGEYGDLLVSGQCEEEEPIQVTEDEVLDILESVLISNMSTSVTRGYALTAIMKLSTRFTCTVNRIKKVVSIYGSSIDVELQQRAVEYNALFKKYDHMRSALLERMPVMEKVTTNGPSEIVQTNGETEPAPLETKPPPSGPQPTSQANDLLDLLGGNDITPVIPTAPTSKPASAGGELLDLLGDITLTGAPAAAPTPASVPQISQPPFLLDGLSSQPLFNDIAPGIPSITAYSKNGLKIEFTFERSNTNPSVTVITIQASNSTELDMTDFVFQAAVPKTFQLQLLSPSSSVVPAFNTGTITQVIKVLNPQKQQLRMRIKLTYNHKGSAMQDLAEVNNFPPQSWQ ap1b1_human AP-1 complex subunit beta-1 Adaptor protein complex AP-1 subunit beta-1 Beta-adaptin 1 Beta-1-adaptin Golgi adaptor HA1/AP1 adaptin beta subunit Clathrin assembly protein complex 1 beta large chain AP1B1 ADTB1 BAM22 CLAPB2 Adaptor-related protein complex 1 subunit beta-1 protein protein human Homo sapiens Human MTDSKYFTTTKKGEIFELKAELNSDKKEKKKEAVKKVIASMTVGKDVSALFPDVVNCMQTDNLELKKLVYLYLMNYAKSQPDMAIMAVNTFVKDCEDPNPLIRALAVRTMGCIRVDKITEYLCEPLRKCLKDEDPYVRKTAAVCVAKLHDINAQLVEDQGFLDTLKDLISDSNPMVVANAVAALSEIAESHPSSNLLDLNPQSINKLLTALNECTEWGQIFILDCLANYMPKDDREAQSICERVTPRLSHANSAVVLSAVKVLMKFMEMLSKDLDYYGTLLKKLAPPLVTLLSAEPELQYVALRNINLIVQKRPEILKHEMKVFFVKYNDPIYVKLEKLDIMIRLASQANIAQVLAELKEYATEVDVDFVRKAVRAIGRCAIKVEQSAERCVSTLLDLIQTKVNYVVQEAIVVIKDIFRKYPNKYESVIATLCENLDSLDEPEARAAMIWIVGEYAERIDNADELLESFLEGFHDESTQVQLQLLTAIVKLFLKKPTETQELVQQVLSLATQDSDNPDLRDRGYIYWRLLSTDPVAAKEVVLAEKPLISEETDLIEPTLLDELICYIGTLASVYHKPPSAFVEGGRGVVHKSLPPRTASSESAESPETAPTGAPPGEQPDVIPAQGDLLGDLLNLDLGPPVSGPPLATSSVQMGAVDLLGGGLDSLMGDEPEGIGGTNFVAPPTAAVPANLGAPIGSGLSDLFDLTSGVGTLSGSYVAPKAVWLPAMKAKGLEISGTFTRQVGSISMDLQLTNKALQVMTDFAIQFNRNSFGLAPATPLQVHAPLSPNQTVEISLPLSTVGSVMKMEPLNNLQVAVKNNIDVFYFSTLYPLHILFVEDGKMDRQMFLATWKDIPNENEAQFQIRDCPLNAEAASSKLQSSNIFTVAKRNVEGQDMLYQSLKLTNGIWVLAELRIQPGNPSCTDLELSLKCRAPEVSQHVYQAYETILKN ap1s3_human AP-1 complex subunit sigma-3 AP1S3 Clathrin assembly protein complex 1 sigma-1C small chain Golgi adaptor HA1/AP1 adaptin sigma-1C subunit Sigma 1C subunit of AP-1 clathrin Sigma-adaptin 1C Sigma1C-adaptin Adaptor protein complex AP-1 subunit sigma-1C Adaptor-related protein complex 1 subunit sigma-1C protein protein human Homo sapiens Human MIHFILLFSRQGKLRLQKWYITLPDKERKKITREIVQIILSRGHRTSSFVDWKELKLVYKRYASLYFCCAIENQDNELLTLEIVHRYVELLDKYFGNVCELDIIFNFEKAYFILDEFIIGGEIQETSKKIAVKAIEDSDMLQEVSTVSQTMGER arf1_human ADP-ribosylation factor 1 ARF1 protein protein human Homo sapiens Human MGNIFANLFKGLFGKKEMRILMVGLDAAGKTTILYKLKLGEIVTTIPTIGFNVETVEYKNISFTVWDVGGQDKIRPLWRHYFQNTQGLIFVVDSNDRERVNEAREELMRMLAEDELRDAVLLVFANKQDLPNAMNAAEITDKLGLHSLRHRNWYIQATCATSGDGLYEGLDWLSNQLRNQK ap1g1_human AP-1 complex subunit gamma-1 Adaptor protein complex AP-1 subunit gamma-1 Golgi adaptor HA1/AP1 adaptin subunit gamma-1 Gamma1-adaptin Clathrin assembly protein complex 1 gamma-1 large chain AP1G1 ADTG CLAPG1 Adaptor-related protein complex 1 subunit gamma-1 protein protein human Homo sapiens Human MPAPIRLRELIRTIRTARTQAEEREMIQKECAAIRSSFREEDNTYRCRNVAKLLYMHMLGYPAHFGQLECLKLIASQKFTDKRIGYLGAMLLLDERQDVHLLMTNCIKNDLNHSTQFVQGLALCTLGCMGSSEMCRDLAGEVEKLLKTSNSYLRKKAALCAVHVIRKVPELMEMFLPATKNLLNEKNHGVLHTSVVLLTEMCERSPDMLAHFRKLVPQLVRILKNLIMSGYSPEHDVSGISDPFLQVRILRLLRILGRNDDDSSEAMNDILAQVATNTETSKNVGNAILYETVLTIMDIKSESGLRVLAINILGRFLLNNDKNIRYVALTSLLKTVQTDHNAVQRHRSTIVDCLKDLDVSIKRRAMELSFALVNGNNIRGMMKELLYFLDSCEPEFKADCASGIFLAAEKYAPSKRWHIDTIMRVLTTAGSYVRDDAVPNLIQLITNSVEMHAYTVQRLYKAILGDYSQQPLVQVAAWCIGEYGDLLVSGQCEEEEPIQVTEDEVLDILESVLISNMSTSVTRGYALTAIMKLSTRFTCTVNRIKKVVSIYGSSIDVELQQRAVEYNALFKKYDHMRSALLERMPVMEKVTTNGPTEIVQTNGETEPAPLETKPPPSGPQPTSQANDLLDLLGGNDITPVIPTAPTSKPSSAGGELLDLLGDINLTGAPAAAPAPASVPQISQPPFLLDGLSSQPLFNDIAAGIPSITAYSKNGLKIEFTFERSNTNPSVTVITIQASNSTELDMTDFVFQAAVPKTFQLQLLSPSSSIVPAFNTGTITQVIKVLNPQKQQLRMRIKLTYNHKGSAMQDLAEVNNFPPQSWQ vamp4_human Vesicle-associated membrane protein 4 VAMP4 protein protein human Homo sapiens Human MPPKFKRHLNDDDVTGSVKSERRNLLEDDSDEEEDFFLRGPSGPRFGPRNDKIKHVQNQVDEVIDVMQENITKVIERGERLDELQDKSESLSDNATAFSNRSKQLRRQMWWRGCKIKAIMALVAAILLLVIIILIVMKYRT ap1g1-arf1-2 1420985 n/a 1420991 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) n/a 1420992 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1420993 unspecified role unspecified role ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1420994 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) n/a 1420995 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false P84077:p.Trp172Asp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false P84077:p.Ile49Asp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false P84077:p.Val53Asp mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P84077:p.Ala136_Ala137delinsProHis mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false P84077:p.[Leu77Asp;His80Asp] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P84077:p.Lys73Asp mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P84077:p.Gln71Leu mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) association association false false false 1420985 Fig. 3A, 3C, 3E, 3F, 5C, Supp. Fig. 2A Presence of the subunits demonstrated by Coomassie staining of the purified complex, however AP1S3 was not shown to be present. It has been included in this complex as it is present in the single polycistronic vector this complex was cloned into, and also because it was shown to be present in the crystal of this complex. Addition of a di-leucine-containing peptide from VAMP4 led to a 4-fold increase in the amount of Arf1 bound to the AP-1 core. Arf1-W172D tightly bound to GST-AP1-A core independent of the VAMP4 peptide. ap1g1-arf1-1 1420985 n/a 1420995 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false P84077:p.Gln71Leu mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1420994 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) n/a 1420992 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1420993 unspecified role unspecified role ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q10567:p.[Ile85Asp;Val88Asp] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1420991 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) association association false false false 1420985 Fig. 3A, 3E, 3F, 5A, 5B, Supp. Fig. 2A Presence of the subunits demonstrated by Coomassie staining of the purified complex, however AP1S3 was not shown to be present. It has been included in this complex as it is present in the single polycistronic vector this complex was cloned into, and also because it was shown to be present in the crystal of this complex. Addition of a di-leucine-containing peptide from VAMP4 led to a 4-fold increase in the amount of Arf1 bound to the AP-1 core. Addition of peptide failed to rescue Arf1 binding to AP-1 complex containing mutated AP1B1. ap1g1-arf1-3 1420985 n/a 1420994 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) n/a 1420993 unspecified role unspecified role ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1420995 unspecified role unspecified role prey prey n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P84077:p.Gln71Leu mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1420991 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) n/a 1420992 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P22892:p.[Leu68Asp;Leu71Glu;Leu102Glu] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P22892:p.Leu102Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false P22892:p.[Leu68Asp;Leu71Glu] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) association association false false false Fig. 3A, 3B, 5A, 5B, Supp. Fig. 2 Presence of the subunits demonstrated by Coomassie staining of the purified complex, however AP1S3 was not shown to be present. It has been included in this complex as it is present in the single polycistronic vector this complex was cloned into, and also because it was shown to be present in the crystal of this complex. Addition of a di-leucine-containing peptide from VAMP4 led to a 4-fold increase in the amount of Arf1 bound to the AP-1 core. The form of the AP-1 complex containing mutated AP1G1 binds weakly to Arf1 in the absence of peptide, this is enhanced 4-fold when peptide is present. ap1b1-ap1m1 1420986 n/a 1420993 unspecified role unspecified role bait bait region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1420991 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 Fig. 4A ap1g1-ap1m1 1420986 n/a 1420991 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 n/a 1420992 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 4B ap1b1-ap1g1-5 1420987 n/a 1420993 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false Q10567:p.[Ile85Asp;Val88Asp] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n/a 1420996 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig. 4C-H ap1g1-ap1m1-1 1420987 n/a 1420992 unspecified role unspecified role neutral component neutral component P22892:p.[Leu68Asp;Leu71Glu;Leu102Glu] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1420991 unspecified role unspecified role neutral component neutral component region egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false Fig. 4 I-N ap1b1-ap1g1-2 1420988 n/a 1420992 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1420993 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1420994 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) n/a 1420991 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) n/a 1420995 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P84077:p.Gln71Leu mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Supp. Table 1 Rworking = 0.2 Rfree = 0.25 7A ap1b1-ap1g1-4 1420989 n/a 1420991 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) n/a 1420992 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1420993 unspecified role unspecified role neutral component neutral component Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1420994 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association false false false Supp. Fig. 3 Melting temperatures Tm of 56–58C were measured and no sign of melting was observed for any of the constructs at the temperatures T = 25C or T = 37C at which the in vitro and biological experiments were carried out. vamp4-ap1m1-1 1420990 n/a 1420997 unspecified role unspecified role bait bait c-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1420992 unspecified role unspecified role ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P22892:p.[Leu68Asp;Leu71Glu;Leu102Glu] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1420994 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) n/a 1420991 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) n/a 1420993 unspecified role unspecified role ancillary ancillary region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q10567:p.[Ile85Asp;Val88Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) association association false false false Fig. 5D Presence of the subunits demonstrated by Coomassie staining of the purified complex, however AP1S3 was not shown to be present. It has been included in this complex as it is present in the single polycistronic vector this complex was cloned into, and also because it was shown to be present in the crystal of this complex. Arf1 enhanced the binding of wild-type AP-1 to VAMP4