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/ Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Cell (0092-8674) 2013 imex curation Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. james.hurley@nih.gov 02-Aug-2013: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2013-JUL-15 by MGT 02-Aug-2013: Contacted by IntAct-Help. Cell (0092-8674) 2013 Only protein-protein interactions imex curation Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Cell (0092-8674) 2013 imex curation Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. james.hurley@nih.gov 02-Aug-2013: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2013-JUL-15 by MGT 02-Aug-2013: Contacted by IntAct-Help. Cell (0092-8674) 2013 Only protein-protein interactions imex curation Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Cell (0092-8674) 2013 imex curation Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. james.hurley@nih.gov 02-Aug-2013: Contacted by IntAct-Help. Only protein-protein interactions 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. The figure legends and mutations entered still do not correspond for Figures 3A and 3C. EBI-6864698:You have entered for Fig 3A several mutants of Arf1 that are not present in Fig 3A (but are present in Fig 3C). I agree the VAMP4 interactions (Fig 5) can be separate - but can Fig 3A/C be added together? I've added all the mutants as mutation increasing/decreasing etc, even though its a multi-protein complex, as its the relationship to the prey they are looking at. As the other molecules can only really be described as ancillary, I have ignored them as far as effects of the different mutants are concerned. james.hurley@nih.gov Accepted 2013-JUL-15 by MGT 02-Aug-2013: Contacted by IntAct-Help. Cell (0092-8674) 2013 Only protein-protein interactions imex curation Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Cell (0092-8674) 2013 imex curation Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. james.hurley@nih.gov 02-Aug-2013: Contacted by IntAct-Help. Only protein-protein interactions 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 Accepted 2013-JUL-15 by MGT 02-Aug-2013: Contacted by IntAct-Help. Cell (0092-8674) 2013 Only protein-protein interactions imex curation Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Cell (0092-8674) 2013 imex curation Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. james.hurley@nih.gov 02-Aug-2013: Contacted by IntAct-Help. 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 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 Accepted 2013-JUL-15 by MGT 02-Aug-2013: Contacted by IntAct-Help. Cell (0092-8674) 2013 Only protein-protein interactions imex curation Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Structural basis for recruitment and activation of the AP-1 clathrin adaptor complex by Arf1. Cell (0092-8674) 2013 imex curation Ren X., Farías G.G., Canagarajah B.J., Bonifacino J.S., Hurley J.H. james.hurley@nih.gov 02-Aug-2013: Contacted by IntAct-Help. Only protein-protein interactions 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. I've added all the mutants as mutation increasing/decreasing etc, even though its a multi-protein complex, as its the relationship to the prey they are looking at. As the other molecules can only really be described as ancillary, I have ignored them as far as effects of the different mutants are concerned. james.hurley@nih.gov Accepted 2013-JUL-15 by MGT 02-Aug-2013: Contacted by IntAct-Help. Cell (0092-8674) 2013 Only protein-protein interactions imex curation vamp4_human Vesicle-associated membrane protein 4 VAMP4 protein protein human Homo sapiens Human MPPKFKRHLNDDDVTGSVKSERRNLLEDDSDEEEDFFLRGPSGPRFGPRNDKIKHVQNQVDEVIDVMQENITKVIERGERLDELQDKSESLSDNATAFSNRSKQLRRQMWWRGCKIKAIMALVAAILLLVIIILIVMKYRT DIP protein O75379 original sequence version: 118 DIP protein O75379 original sequence version: 128 DIP protein O75379 original sequence version: 133 556CC682D26CF91E 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 178755E8EA4E9ABE 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 DIP protein P35585 original sequence version: 99 1213566B3A681B4C 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 42EF40223DFBA5E9 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 15317E4BCD9503EB 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 FE1BA762F9318585 arf1_human ADP-ribosylation factor 1 ARF1 protein protein human Homo sapiens Human MGNIFANLFKGLFGKKEMRILMVGLDAAGKTTILYKLKLGEIVTTIPTIGFNVETVEYKNISFTVWDVGGQDKIRPLWRHYFQNTQGLIFVVDSNDRERVNEAREELMRMLAEDELRDAVLLVFANKQDLPNAMNAAEITDKLGLHSLRHRNWYIQATCATSGDGLYEGLDWLSNQLRNQK DIP protein P84077 original sequence version: 122 DIP protein P84077 original sequence version: 56 DIP protein P84077 original sequence version: 89 AAC773D4A60186B6 ap1b1-ap1g1-2 2 11 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 ecoli Escherichia coli (strain K12) 12 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 Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain I F Wed May 02 19:36:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile488phe' Tue Jun 30 19:28:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile488Phe' ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 13 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 P84077:p.Gln71Leu mutation mutation certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 20:24:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln71leu' Tue Jun 30 18:57:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln71Leu' ecoli Escherichia coli (strain K12) physical association physical association Supp. Table 1 Rworking = 0.2 Rfree = 0.25 7A ap1g1-arf1-2 3 9 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) 11 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 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 ecoli Escherichia coli (strain K12) 12 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 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 Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain I F Wed May 02 17:24:32 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile488phe' Tue Jun 30 17:28:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile488Phe' ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) 13 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 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 P84077:p.Trp172Asp mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain W D Wed May 02 17:58:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp172asp' Tue Jun 30 19:03:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp172Asp' P84077:p.Ile49Asp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain I D Wed May 02 20:19:20 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile49asp' Tue Jun 30 17:32:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile49Asp' P84077:p.Val53Asp mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain V D Wed May 02 14:31:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val53asp' Tue Jun 30 19:41:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val53Asp' P84077:p.Ala136_Ala137delinsProHis mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain AA PH Thu Jul 07 12:17:47 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was ala136pro_ala137his Fri Jul 22 10:56:35 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'ala136pro,ala137his' Wed May 02 15:10:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala_ala136-137pro_his' Tue Jun 30 18:43:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala136_Ala137delinsProHis' P84077:p.[Leu77Asp;His80Asp] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain L D certain certain sequence position certain certain certain sequence position certain H D Wed May 02 17:13:34 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu77asp,his80asp' Tue Jun 30 17:34:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu77Asp;His80Asp]' P84077:p.Lys73Asp mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain K D Wed May 02 17:35:52 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys73asp' Tue Jun 30 20:23:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys73Asp' P84077:p.Gln71Leu mutation mutation certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 19:38:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln71leu' Tue Jun 30 19:46:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln71Leu' ecoli Escherichia coli (strain K12) association association 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 3 13 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 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 P84077:p.Gln71Leu mutation mutation certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 16:15:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln71leu' Tue Jun 30 18:01:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln71Leu' ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) 11 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 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 ecoli Escherichia coli (strain K12) 12 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 Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain I F Wed May 02 14:41:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile488phe' Tue Jun 30 18:15:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile488Phe' 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 Q10567:p.[Ile85Asp;Val88Asp] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain I D certain certain sequence position certain certain certain sequence position certain V D Wed May 02 19:19:48 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile85asp,val88asp' Tue Jun 30 17:11:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile85Asp;Val88Asp]' ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) association association 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. ap1b1-ap1g1-4 4 9 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) 11 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 ecoli Escherichia coli (strain K12) 12 unspecified role unspecified role neutral component neutral component Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain I F Wed May 02 19:57:52 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile488phe' Tue Jun 30 18:48:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile488Phe' region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) physical association physical association 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. ap1b1-ap1m1 1 12 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 9 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 physical association physical association Fig. 4A ap1g1-ap1m1 1 9 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 11 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 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 physical association physical association Fig. 4B ap1b1-ap1g1-5 5 12 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 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 Q10567:p.[Ile85Asp;Val88Asp] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain I D certain certain sequence position certain certain certain sequence position certain V D Wed May 02 17:04:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile85asp,val88asp' Tue Jun 30 17:54:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile85Asp;Val88Asp]' 10 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig. 4C-H ap1g1-ap1m1-1 5 11 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 L D certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain L E Wed May 02 15:01:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu68asp,leu71glu,leu102glu' Tue Jun 30 19:14:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu68Asp;Leu71Glu;Leu102Glu]' region mcherry mcherry fluorescent protein tag mcherry 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 9 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 colocalization colocalization Fig. 4 I-N ap1g1-arf1-3 3 8 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) 12 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 Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain I F Wed May 02 19:06:39 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile488phe' Tue Jun 30 16:53:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile488Phe' 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 ecoli Escherichia coli (strain K12) 13 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P84077:p.Gln71Leu mutation mutation certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 17:02:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln71leu' Tue Jun 30 18:35:10 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln71Leu' ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) 11 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 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 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 L D certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain L E Wed May 02 18:03:31 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu68asp,leu71glu,leu102glu' Tue Jun 30 18:30:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu68Asp;Leu71Glu;Leu102Glu]' P22892:p.Leu102Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain L E Wed May 02 17:22:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu102glu' Tue Jun 30 16:30:34 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu102Glu' P22892:p.[Leu68Asp;Leu71Glu] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain L D certain certain sequence position certain certain certain sequence position certain L E Wed May 02 20:05:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu68asp,leu71glu' Tue Jun 30 17:29:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu68Asp;Leu71Glu]' ecoli Escherichia coli (strain K12) association association 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. vamp4-ap1m1-1 6 7 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 11 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 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 L D certain certain sequence position certain certain certain sequence position certain L E certain certain sequence position certain certain certain sequence position certain L E Wed May 02 15:04:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu68asp,leu71glu,leu102glu' Tue Jun 30 18:30:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Leu68Asp;Leu71Glu;Leu102Glu]' ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role ancillary ancillary ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 12 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 Q10567:p.[Ile85Asp;Val88Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain I D certain certain sequence position certain certain certain sequence position certain V D Wed May 02 15:36:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile85asp,val88asp' Tue Jun 30 16:03:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile85Asp;Val88Asp]' 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 Q10567:p.Ile488Phe mutation mutation certain certain sequence position certain certain certain sequence position certain I F Wed May 02 17:04:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile488phe' Tue Jun 30 17:41:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ile488Phe' ecoli Escherichia coli (strain K12) association association 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