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/ Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Mol. Biol. Cell (1059-1524) 2003 imex curation Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu 27-Oct-2007: Contacted by JYOTI. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu Accepted 2009-OCT-21 by JYOTI. 27-Oct-2007: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2003 jyoti Only protein-protein interactions imex curation Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Mol. Biol. Cell (1059-1524) 2003 imex curation Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu 27-Oct-2007: Contacted by JYOTI. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast crosslink cross-linking study crosslink anti tag western anti tag western blot anti tag western Conibear E., Cleck JN., Stevens TH. Crosslinking agent Dithiobis[succinimidyl propionate. Cross-linked proteins then immunoprecipitatedwith anti-Tlg1 antibody. stevens@molbio.uoregon.edu Accepted 2009-OCT-21 by JYOTI. 27-Oct-2007: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2003 jyoti Only protein-protein interactions imex curation Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Mol. Biol. Cell (1059-1524) 2003 imex curation Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu 27-Oct-2007: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu Accepted 2009-OCT-21 by JYOTI. 27-Oct-2007: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2003 jyoti Only protein-protein interactions imex curation Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Mol. Biol. Cell (1059-1524) 2003 imex curation Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu 27-Oct-2007: Contacted by JYOTI. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu Accepted 2009-OCT-21 by JYOTI. 27-Oct-2007: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2003 jyoti Only protein-protein interactions imex curation Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Mol. Biol. Cell (1059-1524) 2003 imex curation Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu 27-Oct-2007: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro pull down pull down predetermined predetermined participant predetermined Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu Accepted 2009-OCT-21 by JYOTI. 27-Oct-2007: Contacted by JYOTI. Mol. Biol. Cell (1059-1524) 2003 jyoti Only protein-protein interactions imex curation Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Vps51p mediates the association of the GARP (Vps52/53/54) complex with the late Golgi t-SNARE Tlg1p. Mol. Biol. Cell (1059-1524) 2003 imex curation Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu 27-Oct-2007: Contacted by JYOTI. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Conibear E., Cleck JN., Stevens TH. stevens@molbio.uoregon.edu Accepted 2009-OCT-21 by JYOTI. Mol. Biol. Cell (1059-1524) 2003 jyoti Only protein-protein interactions imex curation vps51_yeast Vacuolar protein sorting-associated protein 51 YKR020W Apical bud growth protein 3 VPS51 API3 VPS67 WHI6 protein protein yeast Saccharomyces cerevisiae Baker's yeast MAEQISHKKSLRVSSLNKDRRLLLREFYNLENEPNKGRQEARIGEKASEAHSGEEQVTDVNIDTEANTEKPVKDDELSATEEDLKEGSEDAEEEIKNLPFKRLVQIHNKLLGKETETNNSIKNTIYENYYDLIKVNDLLKEITNANEDQINKLKQTVESLIKEL 7E76264F654AC452 vps53_yeast Vacuolar protein sorting-associated protein 53 J1258 YJL029C VPS53 protein protein yeast Saccharomyces cerevisiae Baker's yeast MLEGTVDYDPLEDITNILFSKESLNNIDELISITRSYKKQLQEDILKEENELKEHPKNSAEIEASLRKVFQDFKETQDVSASTELTISNLTEGISYLDIAKKNLTHSLTLFQNLKILTDSYIQCNELLSQGSFKKMVSPYKIMCSLAENTFISYKSLDEINYLLSSISRLKGDTLSKIKQNYNALFSGGNISEHDTALTMELREGACELLDCDTSTRAQMIDWCLDKLLFEMKEIFRVDDEAGSLENLSRRYIYFKKILNNFNSKFADYFLKDWEMAVRLTTTFYHITHKDLQTLLKREFKDKNPSIDLFMTALQSTLDFEKYIDVRFSKKIKEPKLSSCFEPYLTLWVSHQNQMMEKKFLSYMSEPKYPSNETESLVLPSSADLFRTYRSVLTQTLELIDNNANDSILTSLANFFSRWLQTYSQKILLPLLLPDNIEVQDKLEAAKYTVLLINTADYCATTIDQLEDKLSEFSGNREKLANSFTKTKNIYDDLLAKGTSFLLNRVIPLDLNFVWREFINNDWSNAAIEDYSRYMVTLKSVLKMPALTDASIKQQQEQPSTLAFILSQFNRDVYKWNFLDKVIDIITTNFVSNTIRLLQPVPPFSLAGSKRKFETRTVVNIGEQLLLDLELLKEIFHTLPESVSNDSDLRENTSYKRVKRHADNNIDQLLKFIKLLMAPLDSADDYYETYSKLTNNNPDSAVWSFVLALKGIPWDLALWKKLWSAYNLETDDTDEGSRPDSNRDLFIFKWDKVLLGQFENNLARMQDPNWSKFVRQDLKISPPVMKRIVSTPQIQQQKEEQKKQSLSVKDFVSHSRFFNRGT C75F19FA57157481 tlg2_yeast t-SNARE affecting a late Golgi compartment protein 2 YOL018C TLG2 Syntaxin TLG2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MFRDRTNLFLSYRRTFPHNITFSSGKAPLGDDQDIEMGTYPMMNMSHDISARLTDERKNKHENHSDALPPIFIDIAQDVDDYLLEVRRLSEQLAKVYRKNSLPGFEDKSHDEALIEDLSFKVIQMLQKCYAVMKRLKTIYNSQFVDGKQLSREELIILDNLQKIYAEKIQTESNKFRVLQNNYLKFLNKDDLKPIRNKASAENTLLLDDEEEEAAREKREGLDIEDYSKRTLQRQQQLHDTSAEAYLRERDEEITQLARGVLEVSTIFREMQDLVVDQGTIVDRIDYNLENTVVELKSADKELNKATHYQKRTQKCKVILLLTLCVIALFFFVMLKPHGGGSGGRNNGSNKYNNDDNKTVNNSHDDGSNTHINDEESNLPSIVEVTESENDALDDLL CEBD6FE11278B1F0 CEBD6FE11278B1F0 tlg1_yeast t-SNARE affecting a late Golgi compartment protein 1 Syntaxin TLG1 TLG1 YDR468C D8035.11 protein protein yeast Saccharomyces cerevisiae Baker's yeast MNNSEDPFQQVVKDTKEQLNRINNYITRHNTAGDDDQEEEIQDILKDVEETIVDLDRSIIVMKRDENEDVSGREAQVKNIKQQLDALKLRFDRRIQESTQTTIPLEETVENSTLNTSMAENNDGGMSNPFQEQMLREQDVHLDGIHKTMQNLHIQAQTMGDELENQGQLLDNMDEGMDGVVNKLARGRRQLEWVYEKNKEKYDDCCIGLLIVVLIVLLVLAFIA B33251C18645EF74 B33251C18645EF74 vps54_yeast Vacuolar protein sorting-associated protein 54 VPS54 CGP1 CPF1 genetically-interacting protein 1 Temperature-sensitive clathrin synthetic mutation protein 3 LUV1 RKI1 TCS3 YDR027C PZF889 YD9813.05C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSISETPHNKSQGLQKAAGRPKIVVPEGSPSRNSDSGSFTIEGDTSLNDDLLSISGSVTPRARRSSRLSLDSITPRRSFDSRTLSVANSRSFGFENETHSGSMDFSPLGNNSIYEIVMNTRRKNWLNYPTVADIPQVSLSKNDLDDHWKTHVIEYVKNIKSDYQIFQSTNNIRNMNQMEQLKELREGENMHEESFEANLRQGDAELINSIPDFYFSDKFQLDNPRTFHKVLDAIDLFLTKLDMKRQAERDEAFSELRDRLNDFLDIVETLLVTEISKSSHKFFHALSEVDNIQKRALDTMSELKELAQNIKTIDAENIRKKISHLEMIFKRKNVEKLEQGLLQAKLVLNKTDECKSMYEENKLDNCLELIKSIDYLIKGDDSINEDVQSWTRCWPYKLSNLRTIPALSATREFLTNMKIEIGGKFSLQLSILLIDDLRSFCKSIKPKETLHRIQTGSNDKKQTIFTDNFSSKITELIVRLNRCEELTSAFDLYREKSITELKSIIKIYLPTENAHADNNHDEKHLNNGSTSGSKLSRLIKEQTPAEFQSMLVNIFTHALEALRRLYGHQKLLLDISLNELASVKSPNENQHNMITQLDIRTGINEIIRIIQLRTGKIIAVRRELNLSLRYDYFLKFYAICVIFIQECEVLSGEFLTKYLSNVLASQIKHYANAQSSKNYRNIKKKIDAEEWIPYIVDSSIQSDVNDIVSSIDIDPLSWTTILDMVGGSHDCENGRSEDKEKDEGNETYQGHRKSVVVGDKTFVASSSLLATIEVIKELMVLSINLPSIYLSNFEKLCYDALQYYNSSAMASVTQPGNSLLKTGRNLSIMGESLDCLAEFVIIVQRFYQRLSNSNRDFEPFDASHYTTLLGQFQASSNKIYMANAPPPPV 67E930C34E8CC5AA vps52_yeast Vacuolar protein sorting-associated protein 52 VPS52 YDR484W D8035.27 SAC2 Suppressor of actin mutation protein 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MDVLKEVLSLDQDKFDQLKETSRDKTNETDDPFENYLKDCKFKAPSNKDQSPFAKLKSLQETHSNNEAAINIIIPQLIDYLTEFTNRLSNYTQDLDFIKKKSNELQSLLEYNSTKLAHISPMVNDLMIPPELIDDIIKGKINESWQDNITFIADKEEIYNKYRSNNLDQDNKDAENSAMLAPKDFDKLCQLLDILKNVILERSKRLIISKIKTLRSHNPVPSQRIQNKLLKVQKIFPFIRDNNLSLALELRQAYCYTMKWYYREYFSRYIRSLTILQFQQIDSQFALGNGLSTTSVSGFNNSPSLFFSNYLTTSASNAFYNKLPVTDEKIDKYFQIKKRLNILTQEDNTVMVSQIAENNTTKNYIEIGFKNLNLAILDNCTVEYHFLKDFFAMNGDNFEEINGLLEQIFQPTFDEATTYTQQLIQYNYDIFGVLISIRVANQLQFESERRGIPSMFDSFLNGQLIQLWPRFQQLVDFQCESLRKAAITTNVAKYAGNSSTSNSSPLTSPHELTVQFGKFLSSFLTLAITHKQSIDERSEPLYNSIIRLRNDFETVMTKCSKKTKSPERFLATNYMYLYNNLQQLHLHLNINDSDAQNYNFDSAENVGTKVANDDDNDSSVPLIIRETENHFKTLVEAFTRN F2BF669A72A712C1 F2BF669A72A712C1 vps51-vps52-1 6 12 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Immunoprecipitation of HA-tagged Vps52p under nondenaturing conditions led to the copurification of either myc-tagged Vps51p (Figure 3A, lane 3) or myc-tagged Vps53p (Figure 3A, lane 5) when these were expressed in the same strain. Figs. 3A and 3B Vps51p and Vps52p could be coimmunoprecipitated only when coexpressed in the same strain, not when lysates from 2 strains were mixed, indicating that the two proteins do not appreciably dissociate and rebind during the course of the experiment. vps52-vps53-1 6 12 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Immunoprecipitation of HA-tagged Vps52p under nondenaturing conditions led to the copurification of either myc-tagged Vps51p (Figure 3A, lane 3) or myc-tagged Vps53p (Figure 3A, lane 5) when these were expressed in the same strain. Fig. 3A vps52-vps53-2 1 8 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Vps51p copurifies and colocalizes with the major cellular pool of Vps52/53/54 complex. Fig. 4A vps52-vps53-3 1 8 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Vps51p copurifies and colocalizes with the major cellular pool of Vps52/53/54 complex. Fig. 4A vps52-vps53-4 1 12 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Vps51p copurifies and colocalizes with the major cellular pool of Vps52/53/54 complex. Fig. 4A vps51-vps52-2 4 7 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Vps51p colocalizes with Vps52p and Vps53p by indirect immunofluorescence microscopy. Fig. 4B vps51-vps53-1 4 7 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Vps51p colocalizes with Vps52p and Vps53p by indirect immunofluorescence microscopy. Fig. 4B vps51-tlg1-1 3 10 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Fig. 7A tlg2-tlg1-1 3 9 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association Fig. 7A vps51-vps52-3 2 10 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role neutral component neutral component 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 Under these conditions, both Vps52p and Vps51p copurified with Tlg1p. Fig. 7B vps51-tlg1-2 2 10 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 7B vps51-vps52-4 5 10 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 ecoli Escherichia coli (strain K12) 12 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 yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 7 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast association association Under these conditions, both Vps52p and Vps51p copurified with Tlg1p. Fig. 7B Exact coordinates of N-terminal domain of TLG1 not given by author. vps51-tlg-3 5 10 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association Under these conditions, both Vps52p and Vps51p copurified with Tlg1p. Fig. 7B Exact coordinates of N-terminal domain of TLG1 not given by author.