UniProt UniProt http://www.uniprot.org panic-2003-2 The ARF-like GTPases Arl1p and Arl3p act in a pathway that interacts with vesicle-tethering factors at the Golgi apparatus. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Panic B., Whyte JR., Munro S. GST fusion proteins have been loaded with either GDP or nonhydrolyzable GTP-analog GTP-gamma-S. Extracts of yeast expressing VPS53 fusion protein have been used. A yeast strain lacking VSP52 was used, leading to an overexpression of VSP53 due to GARP complex disassembly. sean@mcr-lmb.cam.ac.uk Curr. Biol. (0960-9822) 2003 panic-2003-1 The ARF-like GTPases Arl1p and Arl3p act in a pathway that interacts with vesicle-tethering factors at the Golgi apparatus. in vitro In vitro pull down pull down antibody detection identification by antibody antibody detection Panic B., Whyte JR., Munro S. GST fusion proteins have been loaded with either GDP or nonhydrolyzable GTP-analog GTP-gamma-S. Extracts of yeast expressing VPS53 fusion protein have been used. A yeast strain lacking VSP52 was used, leading to an overexpression of VSP53 due to GARP complex disassembly. sean@mcr-lmb.cam.ac.uk Curr. Biol. (0960-9822) 2003 panic-2003-3 The ARF-like GTPases Arl1p and Arl3p act in a pathway that interacts with vesicle-tethering factors at the Golgi apparatus. in vitro In vitro pull down pull down fingerprinting peptide massfingerprinting fingerprinting Panic B., Whyte JR., Munro S. GST fusion proteins have been loaded with either GDP or nonhydrolyzable GTP-analog GTP-gamma-S. Extracts of yeast expressing VPS53 fusion protein have been used. A yeast strain lacking VSP52 was used, leading to an overexpression of VSP53 due to GARP complex disassembly. sean@mcr-lmb.cam.ac.uk Curr. Biol. (0960-9822) 2003 panic-2003-4 The ARF-like GTPases Arl1p and Arl3p act in a pathway that interacts with vesicle-tethering factors at the Golgi apparatus. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Panic B., Whyte JR., Munro S. GST fusion proteins have been loaded with either GDP or nonhydrolyzable GTP-analog GTP-gamma-S. Extracts of yeast expressing VPS53 fusion protein have been used. A yeast strain lacking VSP52 was used, leading to an overexpression of VSP53 due to GARP complex disassembly. sean@mcr-lmb.cam.ac.uk Curr. Biol. (0960-9822) 2003 panic-2003-5 The ARF-like GTPases Arl1p and Arl3p act in a pathway that interacts with vesicle-tethering factors at the Golgi apparatus. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast confocal microscopy confocal microscopy anti tag immunost anti tag immunostaining anti tag immunost Panic B., Whyte JR., Munro S. GST fusion proteins have been loaded with either GDP or nonhydrolyzable GTP-analog GTP-gamma-S. Extracts of yeast expressing VPS53 fusion protein have been used. A yeast strain lacking VSP52 was used, leading to an overexpression of VSP53 due to GARP complex disassembly. sean@mcr-lmb.cam.ac.uk Curr. Biol. (0960-9822) 2003 imh1_yeast Golgin IMH1 Integrins and myosins homology protein 1 YLR309C IMH1 SYS3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MFKQLSQIGKNLTDELAKGLADDMSPTPSEQQIEDDKSGLPKEIQAKLRKFEKYEQKYPLLLSAYKNEKLKSEKLEAVEKILAENTPISNIDDAVDTLPAFFQDLNNKNNLLNDEIKRLTKQNSEIPESASSETLKDKEEEFLKKEQNYKNDIDDLKKKMEALNIELDTVQKEKNDTVSGLREKIVALENILKEEREAKKQKEEVSISELKEELAIKNHSLEDSRMKITELEQNLSSKSTIMEEKSSELAELNITLKEKERKLSELEKKMKELPKAISHQNVGNNNRRKKNRNKGKKNKGGITTGDISEEETVDNSINTEEYDKLKENLQELQEKYKDCEDWKQKYEDIEAELKDAKELENSQLEKSAKELETLNTELIDTKKSLKEKNSELEEVRDMLRTVGNELVDAKDEIKESSSKQNEEVKTVKLELDDLRHKNATMIEAYEAKNTELRSKIELLSKKVEHLKNLCTEKEKEQTTSQNKVAKLNEEISQLTYEKSNITKELTSLRTSYKQKEKTVSYLEEQVKQFSEQKDVAEKSTEQLRKDHAKISNRLDLLKKENETLHNDIAKNSNSYEEYLKENGKLSERLNILQEKYNTLQNVKSNSNEHIDSIKRQCEELNVKLKESTKKILSLEDELNEYANIVQDKTREANTLRRLVSDSQTDDSSKQKELENKLAYLTDEKNKLEAELDLQTSRKATELQEWKHTVTELKSEIHALKLREEGLKSEVDALKHVNNDIKRKTQATSDDSDQLEQITSNLKLSLSKADEKNFELQSANEKLLNLNNELNKKFDRLLKNYRSLSSQLNALKERQYSDKSGRVSRSGSIGTLANANIDSSPANNSNPTKLEKIRSSSSLELDSEKNEKIAYIKNVLLGFLEHKEQRNQLLPVISMLLQLDSTDEKRLVMSLK 090BD9AA5702C01D arl1_yeast ADP-ribosylation factor-like protein 1 ARL1 Arf-like GTPase 1 ARF3 YBR1216 YBR164C protein protein yeast Saccharomyces cerevisiae Baker's yeast MGNIFSSMFDKLWGSNKELRILILGLDGAGKTTILYRLQIGEVVTTKPTIGFNVETLSYKNLKLNVWDLGGQTSIRPYWRCYYADTAAVIFVVDSTDKDRMSTASKELHLMLQEEELQDAALLVFANKQDQPGALSASEVSKELNLVELKDRSWSIVASSAIKGEGITEGLDWLIDVIKEEQL slo1_yeast SCOCO-like protein 1 SLO1 YER180C-A protein protein yeast Saccharomyces cerevisiae Baker's yeast MSAENISTGSPTGKQPSSEVNLGEREAGTKNERMMRQTKLLKDTLDLLWNKTLEQQEVCEQLKQENDYLEDYIGNLMRSSNVLEK vps53_yeast Vacuolar protein sorting-associated protein 53 J1258 YJL029C VPS53 protein protein yeast Saccharomyces cerevisiae Baker's yeast MLEGTVDYDPLEDITNILFSKESLNNIDELISITRSYKKQLQEDILKEENELKEHPKNSAEIEASLRKVFQDFKETQDVSASTELTISNLTEGISYLDIAKKNLTHSLTLFQNLKILTDSYIQCNELLSQGSFKKMVSPYKIMCSLAENTFISYKSLDEINYLLSSISRLKGDTLSKIKQNYNALFSGGNISEHDTALTMELREGACELLDCDTSTRAQMIDWCLDKLLFEMKEIFRVDDEAGSLENLSRRYIYFKKILNNFNSKFADYFLKDWEMAVRLTTTFYHITHKDLQTLLKREFKDKNPSIDLFMTALQSTLDFEKYIDVRFSKKIKEPKLSSCFEPYLTLWVSHQNQMMEKKFLSYMSEPKYPSNETESLVLPSSADLFRTYRSVLTQTLELIDNNANDSILTSLANFFSRWLQTYSQKILLPLLLPDNIEVQDKLEAAKYTVLLINTADYCATTIDQLEDKLSEFSGNREKLANSFTKTKNIYDDLLAKGTSFLLNRVIPLDLNFVWREFINNDWSNAAIEDYSRYMVTLKSVLKMPALTDASIKQQQEQPSTLAFILSQFNRDVYKWNFLDKVIDIITTNFVSNTIRLLQPVPPFSLAGSKRKFETRTVVNIGEQLLLDLELLKEIFHTLPESVSNDSDLRENTSYKRVKRHADNNIDQLLKFIKLLMAPLDSADDYYETYSKLTNNNPDSAVWSFVLALKGIPWDLALWKKLWSAYNLETDDTDEGSRPDSNRDLFIFKWDKVLLGQFENNLARMQDPNWSKFVRQDLKISPPVMKRIVSTPQIQQQKEEQKKQSLSVKDFVSHSRFFNRGT 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 arl1-imh1-2 665598 N/A 665603 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 tyr780ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 665604 unspecified role unspecified role bait bait region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 ecoli Escherichia coli (strain K12) physical association physical association false false false By GTP. 2B, C and D arl3-slo1-1 665598 N/A 665605 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 Q3E784:p.Tyr72Ala mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 665604 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) physical association physical association false false false 3C and D Interaction was independent of ARL1 nucleotide state (GDP / GTP). arl1-vps53-1 665598 N/A 665606 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 665604 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) physical association physical association false false false 4B By GTP. arl1-vps54-1 665598 N/A 665604 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) N/A 665607 unspecified role unspecified role prey prey region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 4B By GTP. arl1-imh1-1 665599 N/A 665604 unspecified role unspecified role bait bait region binding region binding-associated region binding region 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 ecoli Escherichia coli (strain K12) N/A 665603 unspecified role unspecified role prey prey physical association physical association false false false By GTP. 2A arl1-vps54-3 665600 N/A 665606 unspecified role unspecified role prey prey N/A 665607 unspecified role unspecified role prey prey N/A 665604 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) association association false false false 4A By GTP. arl1-vps53-2 665601 N/A 665606 unspecified role unspecified role prey prey region protein a tag protein a tag protein A protein A undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 665604 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) physical association physical association false false false The interaction is independent of GDP and GTP. 4C arl1-vps54-2 665602 N/A 665604 unspecified role unspecified role neutral component neutral component region rfp tag red fluorescent protein tag red fluorescent protein RFP red fluorescent protein RFP rfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 665607 unspecified role unspecified role neutral component neutral component region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false colocalization colocalization false false false 4D