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/ Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Neuron (0896-6273) 2000 Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library davet rat-hippocampus Rattus norvegicus hippocampal neurons hippocampus Hippocampus [Brain hippocampus] confocal microscopy confocal microscopy immunostaining immunostaining Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu Accepted 2009-NOV-16 by DAVET. 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library Neuron (0896-6273) 2000 davet Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Neuron (0896-6273) 2000 Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library davet 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu Accepted 2009-NOV-16 by DAVET. 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library Neuron (0896-6273) 2000 davet Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Neuron (0896-6273) 2000 Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library davet 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 Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu Accepted 2009-NOV-16 by DAVET. 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library Neuron (0896-6273) 2000 davet Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Neuron (0896-6273) 2000 Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library davet in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa antibody detection identification by antibody antibody detection Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu Accepted 2009-NOV-16 by DAVET. 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library Neuron (0896-6273) 2000 davet Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Neuron (0896-6273) 2000 Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library davet in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting anti tag western anti tag western blot anti tag western Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu Accepted 2009-NOV-16 by DAVET. 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library Neuron (0896-6273) 2000 davet Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Neuron (0896-6273) 2000 Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library davet rat-brain Rattus norvegicus brain brain Brain [cerebrum] density sedimentation cosedimentation through density gradient density sedimentation western blot western blot Immuno blot Immuno blot Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu Accepted 2009-NOV-16 by DAVET. 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library Neuron (0896-6273) 2000 davet Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Piccolo, a presynaptic zinc finger protein structurally related to bassoon. Neuron (0896-6273) 2000 Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library davet rat-brain Rattus norvegicus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Fenster SD., Chung WJ., Zhai R., Cases-Langhoff C., Voss B., Garner AM., Kaempf U., Kindler S., Gundelfinger ED., Garner CC. Synapse - Interactions of proteins with an established role in the presynapse. The Bait assumed to be of rat in origin since this is the only Piccolo cDNA discussed by the authors. The authors have however, not specifically mentioned the origin of the protien used. no Hippocampal neurons cultured for 24 days were studied. Synaptotagmin and GABAA are distributed over multiple uniprot entries. These were also shown to colocalize with Piccolo protein. garner@nrc.uab.edu Accepted 2009-NOV-16 by DAVET. 23-Dec-2009: Contacted by JYOTI. Mouse brain MATCHMAKER LexA cDNA library Neuron (0896-6273) 2000 davet praf1_mouse Prenylated Rab acceptor protein 1 PRA1 family protein 1 Prenylin Rabac1 Pra Pra1 Praf1 protein protein mouse Mus musculus Mouse MAAQKDQQKDAEGEGLSATTLLPKLIPSGAGREWLERRRATIRPWGTFVDQQRFSRPRNVGELCQRLVRNVEYYQSNYVFVFLGLILYCVVTSPMLLVALAVFFGACYILYLRTLQSKLVLFGREVSPAHQYALAGGVSFPFFWLAGAGSAVFWVLGATLVLIGSHAAFHQMEPADGEELQMEPV 86C82C0502B5CB5B pclo_rat Protein piccolo Pclo Aczonin Multidomain presynaptic cytomatrix protein protein protein rat Rattus norvegicus (Rat) Rat MGNEASLEGEGLPEGLAAAAGAGGSGSALHPGIPAGMEADLSQLSEEERRQIAAVMSRAQGLPKGSVPPAAAESPSMHRKQELDSSQAPQQPGKPPDPGRPTQPGLSKSRTTDTFRSEQKLPGRSPSTISLKESKSRTDFKEEYKSSMMPGFFSDVNPLSAVSSVVNKFNPFDLISDSEASQEETTKKQKVVQKEQGKSEGMAKPPLQQPSPKPIPKQQGQVKEVIQQDSSPKSVSSQQAEKVKPQAPGTGKPSQQSPAQTPAQQASPGKPVAQQPGSAKATVQQPGPAKSPAQPAGTGKSPAQPPAKTPGQQAGLEKTSSSQQPGPKSLAQTPGHGKFPLGPVKSPAQQPGTAKHPAQQPGPQTAAKVPGPTKTPAQQSGPGKTPAQQPGPTKPSPQQPIPAKPQPQQPVATKTQPQQSAPAKPQPQQPAPAKPQPQQPTPAKPQPQPPTPAKPQPQPPTATKPQPQPPTATKPHHQQPGLAKPSAQQPTKSISQTVTGRPLQPPPTSAAQTPAQGLSKTICPLCNTTELLLHIPEKANFNTCTECQSTVCSLCGFNPNPHLTEIKEWLCLNCQMQRALGGDLAAAIPSSPQPTPKAATAPTATASKSPVPSQQASPKKEPPSKQDSPKALESKKPPEPKKPPEPKKPPEPKKPPPLVKQPTLHGPTPATAPQLPVAEALPEPAPPKEPSGPLPEQAKAPVGDVEPKQPKMTETRADIQSSSTTKPDILSSQVQSQAQVKTASPLKTDSAKPSQSFPPTGEKTTPLDSKAMPRPASDSKIISQPGPGSESKDPKHIDPIQKKDEPKKAQPKGSPKPETKPVPKGSPTPSGTRPTAGQAAPPSQQPPKPQEQSRRFSLNLGGITDAPKSQPTTPQETVTGKLFGFGASIFSQASNLISTAGQQGPHPQTGPAAPSKQAPTPSQSPAAQGPAKSTGQLPPAPAKATAVKKEAKAAAAENLESKPEQAPTAKKTEKDKKPPPAKVGKPPPSEPEKAVPAHKPDKTTKPKPACPLCRTELNLGSQEPPNFNTCTECKNQVCNLCGFNPTPHLTEIQEWLCLNCQTQRAISGQLGDMGKMPPAPSGPKASPMPAPAEPSSQKTPTGTQVKGKKKEAEGKTEAEKPVPEKETASIEKTPPMVTTDQKLEESEGKKSKVSALPEKKPSEEEKAISADKKERKPPAEEKPPLEEKKPIPVDKKLPPEAKPLSSEGEEKHEILKAHVQIPEEEPTGKVAAKAGEEEQQPDSRPEALPGATPLTLPKAGEKERAVAQPQAEGSSKDGQGERSKEKTEKEEDKSDTSSSQQPKSPQGLSDTGYSSDGISGSLGEIPSLIPSDEKDLLKGLKKDSFSQESSPSSPSDLAKLESTVLSILEAQASTLVGEKAEKKTQPQKISPEKPQDQQKTQTASETLDITISEEEIKESQEKKVSPKKDSEQGFPSRKEHKEKPELVDDLSPRRASYDSVEDSSESENSPVVRRKRRTSIGSSSSDEYKQEDSQGSGEEEDFIRKQIIEMSADEDASGSEDEEFIRSQLKEISGVGESQKREEAKGKGKGVAGKHRRLTRKSSTSFDDDAGRRHSWHDEDDETFDESPELKFRETKSQESEELVVAGGGGLRRFKTIELNSTIADKYSSESSQKKTILYFDEEPELEMESLTDSPEDRSRGEGSSSLHASSFTPGTSPTSVSSLDEDSDSSPSHKKGESKQQRKARHRSHGPLLPTIEDSSEEEELREEEELLKEQEKQRELEQQQRKSSSKKSKKDKDELRAQRRRERPKTPPSNLSPIEDASPTEELRQAAEMEELHRSSCSEYSPSIESDPEGFEISPEKIIEVQKVYKLPAAVSLYSPTDEQSVMQKEGVQKALKSAEEMYEEMMQKPHKYKAFPAANERDEVFEKEPLYGGMLIEDYIYESLVEDTYNGSVDGSLLTRQEEQNGFMQQRGREQKVRLQEQIYDDPMQKISDLQKEFYELESLHSVVPQEDIVSSSYIIPESHEIVDLGSMVMSTSEEKKLLDADSAYEELMRRQQVQVTDGSSPVQTTIGDDMAESTLDFDRVQDASLTSSILSGASLTDSTSSATLSIPDVKITQQFSAEELEDEYVTDYTREIQDIIAHESLILTYSEPSESATSVPPSDTPSLTSSISSVCTTDSSSPVTTLDSLTTVYTEPADVMTKFKDSEEISSTYFPGSIIDYPEDISVSLDRTIMPESRTNEDRIVLSFSGMAPSVVESVGTKPERPQADTISTDLPISEKDLIKGKKETGDGIILEVLDAYKDKREESEAELTKISLPEPGLAQAPSSVTAPQIKEQHVSPHSVSGKISGQEKPTYRLPSGSLPVSTHPSKSRPFFRSSSLDISAQPPPPPPPPPPSPSTSSPPPTPPLPPATSPKPPTYPKKKLAVAATVTSTTIVTTHVDALTMVEAAAARRSNGLPATKMCAIAPPPVPPKPSQIPTGLVFTHRPEAIKPPIAPKPAVPQIPVTTQKPTDTCPKPTGLSLTSTMSLNLVTSADYNVPSPTSPLSPHSNKSSPRYSKSLMDTYVVITLPSEPGTPTDSSAAQAITSWPLGSPPKDLVSLETVFSVVPPMTSTEIPSASQPTLYTSGALGTFSVTPAVTASLFQTVPTSLTQFLPAEASKPEVSAVSSAVPSVAPRSVSIPIPPEPLALDRHQYKENGKLPLIGDAIDLRTIPKSEVKVTEKCMDLSASAMDVKRQTTANEVYRRQISAVQPSIINLSAASSLGTPVTMDSKTVAVVTCTDTTIYTTGTESQVGIEHAVTSPLQLTTSKHTELPYRKPSSQAFPTIRDEAPINLSLGPSAQAVTLAVTKPVTVPPVGVTNGWTDSTLSQGVADGEVVDLSTSKSHRTVVTMDESTSNVVTKIIEDDEKPVDLTAGRRAVCCDMVYTLPFGRSCTAQQPATTLPEDRFGYRDDHYQYDRSGPYGYRGIGGMKPSMSDTNLPEAGHFFYKSKNAFDYSGGTGAAVDLTSGRVSTGEVMDYSSKTTGPYPETRQVISGVGISTPQYSTARLTPPPGPQYGVGSVLRSSNGVVYSSVATPIPSTFAITTQPGSIFSTTVRDLSGIPTTDAMTSLSALHQSQPMPRSYFITTGASETDIAVTGIDINASLQTITMETLPAETMDSVPTLTTASEVFSEVVGEESTLLIVPDEDKQQQQLDLERELLELEKIKQQRFAEELEWERQEIQRFREQEKIMVQKKLEELQSMKQHLLYQQEEERQAQFMMRQETLAQQQLQLEQIQQLQQQLHQQLEEQKLRQIYQYNYDPSGTSSPQTTTEQAILEGQYAATEGSQFWATEDATTTASTVVAIEIPQSQGWYTVQSDGVTQYIAPPGILSTVSEIPLTDVVVKEEKQPKKRSSGAKVRGQYDEMGESVADDPRNLKKIVDSGVQTDDEETADRSYASRRRRTKKSVDTSVQTDDEDQDEWDMPSRSRRKARTGKYGDSTAEGDKTKPLSKVSSVAVQTVAEISVQTEPVGTIRTPSIRARVDAKVEIIKHISAPEKTYKGGSLGCQTETDSDTQSPPYLGATSPPKDKKRPTPLEIGYSSSHLRADPTVQLAPSPPKSPKVLYSPISPLSPGNALEPAFVPYEKPLPDDISPQKVLHPDMAKVPPASPKTAKMMQRSMSDPKPLSPTADESSRAPFQYSEGFTTKGSQTMTASGTQKKVKRTLPNPPPEEVSTGTQSTYSTMGTASRRRMCRTNTMARAKILQDIDRELDLVERESAKLRKKQAELDEEEKEIDAKLRYLEMGINRRKEALLKEREKRERAYLQGVAEDRDYMSDSEVSSTRPSRVESQHGVERPRTAPQTEFSQFIPPQTQTEAQLVPPTSPYTQYQYSSPALPTQAPTPYTQQSHFQQQTLYHQQVSPYQTQPTFQAVATMSFTPQAQPTPTPQPSYQLPSQMMVIQQKPRQTTLYLEPKITSNYEVIRNQPLMIAPVSTDNTYAVSHLGSKYNSLDLRIGLEERSSMAGSPISSISADSFYADIDHHTSRNYVLIDDIGDITKGTAALSTVFSLHEKDLSKTDRLLRTTETRRSQEVTDFLAPLQTSSRLHSYVKADEDPMEDPYELKLLKHQIKQEFRRGTESLDHLAGLSHYYHADTSYRHFPKSEKYSISRLTLEKQAAKQLPAAILYQKQSKHKKSLIDPKMSKFSPIQESRDLEPDYPTYMSSGTSSIGGISSRARLLQDDITFGLRKNITDQQKFMGSSLGSGLGTLGNTIRSALQDEADKPYSSGSRSRPSSRPSSVYGLDLSIKRDSSSSSLRLKAQEAEALDVSFGHSSSSARTKPTSLPISQSRGRIPIVAQSSEEESPLSPVGQPMGMARAAAGPLPPISADTRDQFGSSHSLPEVQQHMREESRTRGYDRDIAFIMDDFQHAMSDSEAYHLRREETDWFDKPRESRLENGHGLDRKLPERLVHSRPLSQHQEQILQMNGKTIHYIFPHARVKITRDFKDHTGSGNGLGIRIVGGKEIPGHSGEIGAYIAKILPGESAEHTGPLMEGMQVLEWNGVPLTSKTYEEVQSIINQQSGEAEICVRLDLNMLSDSENPQHLELHEPPKVDKAKSPGVDPKQLAAELQKVSLQQSPLVMSSVVEKGSHAHSGPTSAGSSSVPSPGQPGSPSVSKKKHSSTKPTDGPKAASHPITGEIQLQINYDLGNLIIHILQARNLVPRDNNGYSDPFVKVYLLPGRGQVMVVQNASAEYKRRTKYVQKSLNPEWNQTVIYKSISMEQLMKKTLEVTVWDYDRFSSNDFLGEVLIDLSSTSHLDNTPRWYPLKEQTESIDHGKSHSSQNSQQSPKPSVIKSRSHGIFPDPSKDMQVPTIEKSHSSPGSSKSSSEGHLRSHGPSRSQSKTSVAQTHLEDAGVAIAAAEAAVQQLRIQPTKPTNHRPAESSVSTGSSGSSVGSGYSVDSEGSSCVAGEPNLLPIPRIGKMGQNGQDPVKQPGMGATDTEGKTQVMGEIKLALKKEMKTDGEQLIVEILQCRNITYKFKSPDHLPDLYVKLYVINISTQKKVIKKKTRVCRHDREPSFNETFRFSLSPAGHSLQILLFSNGGKFMKKTLIGEACIWLDKVDLRKRIVNWHKLLVSPTQTH 5A1BB543201A7450 praf1_rat Prenylated Rab acceptor protein 1 Rabac1 Pra1 Praf1 PRA1 family protein 1 protein protein rat Rattus norvegicus (Rat) Rat MAAQKDQQKDAEVEGLSATTLLPKLIPSGAGREWLERRRATIRPWGTFVDQQRFSRPRNVGELCQRLVRNVEYYQSNYVFVFLGLILYCVVTSPMLLVALAVFFGACYILYLRTLQSKLVLFGREVSPAHQYALAGGVSFPFFWLAGAGSAVFWVLGATLVLIGSHAAFHQIEPADGEELQMEPV DA6341AE66F5C2F0 rab3a_rat Ras-related protein Rab-3A Rab3a protein protein rat Rattus norvegicus (Rat) Rat MASATDSRYGQKESSDQNFDYMFKILIIGNSSVGKTSFLFRYADDSFTPAFVSTVGIDFKVKTIYRNDKRIKLQIWDTAGQERYRTITTAYYRGAMGFILMYDITNEESFNAVQDWSTQIKTYSWDNAQVLLVGNKCDMEDERVVSSERGRQLADHLGFEFFEASAKDNINVKQTFERLVDVICEKMSESLDTADPAVTGAKQGPQLTDQQAPPHQDCAC 1A3E9F8C9D09EB40 rp3a_mouse Rabphilin-3A Rph3a Exophilin-1 protein protein mouse Mus musculus Mouse MTDTVVNRWMYPGDGPLQSNDKEQLQAGWSVHPGAQTDRQRKQEELTDEEKEIINRVIARAEKMEAMEQERIGRLVDRLETMRKNVAGDGVNRCILCGEQLGMLGSACVVCEDCKKNVCTKCGVETSNNRPHPVWLCKICLEQREVWKRSGAWFFKGFPKQVLPQPMPIKKTKPQQPAGEPATQEQPTPESRHPARAPARGDMEDRRPPGQKPGPDLTSAPGRGSHGPPTRRASEARMSTAARDSEGWDHAHGGGTGDTSRSPAGLRRANSVQAARPAPAPVPSPAPPQPVQPGPPGGSRATPGPGRFPEQSTEAPPSDPGYPGAVAPAREERTGPAGGFQAAPHTAAPYSQAAPARQPPPAEEEEEEANSYDSDEATTLGALEFSLLYDQDNSNLQCTIIRAKGLKPMDSNGLADPYVKLHLLPGASKSNKLRTKTLRNTRNPVWNETLQYHGITEEDMQRKTLRISVCDEDKFGHNEFIGETRFSLKKLKANQRKNFNICLERVIPMKRAGTTGSARGMALYEEEQVERIGDIEERGKILVSLMYSTQQGGLIVGIIRCVHLAAMDANGYSDPFVKLWLKPDMGKKAKHKTQIKKKTLNPEFNEEFFYDIKHSDLAKKSLDISVWDYDIGKSNDYIGGCQLGISAKGERLKHWYECLKNKDKKIERWHQLQNENHVSSD D09F8D8D2CBB271E vamp2_rat Vesicle-associated membrane protein 2 Vamp2 Syb2 Synaptobrevin-2 protein protein rat Rattus norvegicus (Rat) Rat MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDIMRVNVDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLKMMIILGVICAIILIIIIVYFST DIP protein P63045 original sequence version: 95 DIP protein P63045 original sequence version: 90 DIP protein P63045 original sequence version: 64 DIP protein P63045 original sequence version: 62 DIP protein P63045 original sequence version: 85 DIP protein P63045 original sequence version: 104 DIP protein P63045 original sequence version: 77 DIP protein P63045 original sequence version: 75 DIP protein P63045 original sequence version: 106 4A0D0D56B5409D0A syph_rat Synaptophysin Major synaptic vesicle protein p38 Syp protein protein rat Rattus norvegicus (Rat) Rat MDVVNQLVAGGQFRVVKEPLGFVKVLQWVFAIFAFATCGSYTGELRLSVECANKTESALNIEVEFEYPFRLHQVYFDAPSCVKGGTTKIFLVGDYSSSAEFFVTVAVFAFLYSMGALATYIFLQNKYRENNKGPMMDFLATAVFAFMWLVSSSAWAKGLSDVKMATDPENIIKEMPMCRQTGNTCKELRDPVTSGLNTSVVFGFLNLVLWVGNLWFVFKETGWAAPFMRAPPGAPEKQPAPGDAYGDAGYGQGPGGYGPQDSYGPQGGYQPDYGQPASGGGGYGPQGDYGQQGYGQQGAPTSFSNQM A245E4F70BF0EAF0 rab3a_mouse Ras-related protein Rab-3A Rab3a protein protein mouse Mus musculus Mouse MASATDSRYGQKESSDQNFDYMFKILIIGNSSVGKTSFLFRYADDSFTPAFVSTVGIDFKVKTIYRNDKRIKLQIWDTAGQERYRTITTAYYRGAMGFILMYDITNEESFNAVQDWSTQIKTYSWDNAQVLLVGNKCDMEDERVVSSERGRQLADHLGFEFFEASAKDNINVKQTFERLVDVICEKMSESLDTADPAVTGAKQGPQLTDQQAPPHQDCAC DIP protein P63011 original sequence version: 100 DIP protein P63011 original sequence version: 53 1A3E9F8C9D09EB40 gria1_rat Glutamate receptor 1 Gria1 GluR-A GluR-K1 Glutamate receptor ionotropic, AMPA 1 AMPA-selective glutamate receptor 1 Glur1 protein protein rat Rattus norvegicus (Rat) Rat MPYIFAFFCTGFLGAVVGANFPNNIQIGGLFPNQQSQEHAAFRFALSQLTEPPKLLPQIDIVNISDSFEMTYRFCSQFSKGVYAIFGFYERRTVNMLTSFCGALHVCFITPSFPVDTSNQFVLQLRPELQEALISIIDHYKWQTFVYIYDADRGLSVLQRVLDTAAEKNWQVTAVNILTTTEEGYRMLFQDLEKKKERLVVVDCESERLNAILGQIVKLEKNGIGYHYILANLGFMDIDLNKFKESGANVTGFQLVNYTDTIPARIMQQWRTSDSRDHTRVDWKRPKYTSALTYDGVKVMAEAFQSLRRQRIDISRRGNAGDCLANPAVPWGQGIDIQRALQQVRFEGLTGNVQFNEKGRRTNYTLHVIEMKHDGIRKIGYWNEDDKFVPAATDAQAGGDNSSVQNRTYIVTTILEDPYVMLKKNANQFEGNDRYEGYCVELAAEIAKHVGYSYRLEIVSDGKYGARDPDTKAWNGMVGELVYGRADVAVAPLTITLVREEVIDFSKPFMSLGISIMIKKPQKSKPGVFSFLDPLAYEIWMCIVFAYIGVSVVLFLVSRFSPYEWHSEEFEEGRDQTTSDQSNEFGIFNSLWFSLGAFMQQGCDISPRSLSGRIVGGVWWFFTLIIISSYTANLAAFLTVERMVSPIESAEDLAKQTEIAYGTLEAGSTKEFFRRSKIAVFEKMWTYMKSAEPSVFVRTTEEGMIRVRKSKGKYAYLLESTMNEYIEQRKPCDTMKVGGNLDSKGYGIATPKGSALRNPVNLAVLKLNEQGLLDKLKNKWWYDKGECGSGGGDSKDKTSALSLSNVAGVFYILIGGLGLAMLVALIEFCYKSRSESKRMKGFCLIPQQSINEAIRTSTLPRNSGAGASGGGGSGENGRVVSQDFPKSMQSIPCMSHSSGMPLGATGL DIP protein P19490 original sequence version: 146 DIP protein P19490 original sequence version: 142 DIP protein P19490 original sequence version: 117 DIP protein P19490 original sequence version: 127 DIP protein P19490 original sequence version: 131 DIP protein P19490 original sequence version: 129 2D4CFA7CCD532838 praf1_human Prenylated Rab acceptor protein 1 PRA1 family protein 1 RABAC1 PRA1 PRAF1 protein protein human Homo sapiens Human MAAQKDQQKDAEAEGLSGTTLLPKLIPSGAGREWLERRRATIRPWSTFVDQQRFSRPRNLGELCQRLVRNVEYYQSNYVFVFLGLILYCVVTSPMLLVALAVFFGACYILYLRTLESKLVLFGREVSPAHQYALAGGISFPFFWLAGAGSAVFWVLGATLVVIGSHAAFHQIEAVDGEELQMEPV A85BD5BDCF6C23E4 bsn_rat Protein bassoon Bsn protein protein rat Rattus norvegicus (Rat) Rat MGNEASLEGGAGEGPLPPGGSGLGPGPGAGKPPSALAGGGQLPVAGAARAAGPPTPGLGLVPGPGPGPGPGSVSRRLDPKEPLGSQRATSPTPKQASATAPGRESPRETRAQGLSGQEAEGPRRTLQVDSRTQRSGRSPSVSPDRGSTPTSPYSVPQIAPLPSSTLCPICKTSDLTSTSSQPNFNTCTQCHNKVCNQCGFNPNPHLTQVKEWLCLNCQMQRALGMDMTTAPRSKSQQQLHSPALSPAHSPAKQPLGKPEQERSRSPGATQSGPRQAEAARATSVPGPTQATAPPEVGRVSPQPPLSTKPSTAEPRPPAGEAQGKSATTVPSGLGAAEQTQGGLTGKLFGLGASLLTQASTLMSVQPEADTQGQPSPSKGPPKIVFSDASKEAGPRPPGSGPGPGPTPGAKTEPGPRTGPGSGPGALAKTGGTPSPKHGRADHQAASKAAAKPKTMPKERAACPLCQAELNVGSRGPANYNTCTACKLRVCTLCGFNPTPHLVEKTEWLCLNCQTKRLLEGSLGEPAPLPLPTPQEPPAGVPQRAAGASPLKQKGPQGPGQPSGSLPPKASPQAAKASPQAAKASPQAKPLRASEPSKTSSSAPEKKTGIPVKAEPVPKPPPETAVPPGTPKAKSGVKRTDPATPVVKPVPEAPKSGEAEEPVPKPYSQDLSRSPQSLSDTGYSSDGVSSSQSEITGVVQQEVEQLDSAGVTGPRPPSPSELHKVGSSMRPSLEAQAVAPSGEWSKPPSGSAVEDQKRRPHSLSIMPEAFDSDEELGDILEEDDSLAMGRQREQQDTAESSDDFGSQLRHDYVEDSSEGGLSPLPPQPPARADMTDEEFMRRQILEMSAEEDNLEEDDTAVSGRGLAKHGAQKASARPRPESSQESVALPKRRLPHNATTGYEELLSEEGPAEPTDGALQGGLRRFKTIGLNSTGRLWSTSLDLGQGSDPNLDREPELEMESLTGSPEDRSRGEHSSTLPASTPSYTSGTSPTSLSSLEEDSDSSPSRRQRLEEAKQQRKARHRSHGPLLPTIEDSSEEEELREEEELLREQEKMREVEQQRIRSTARKTRRDKEELRAQRRRERSKTPPSNLSPIEDASPTEELRQAAEMEELHRSSCSEYSPSPSLDSEAETLDGGPTRLYKSGSEYNLPAFMSLCSPTETPSGSSTTPSSGRPLKSAEEAYEDMMRKAELLQRQQGQAAGARGPHGGPSQPTGPRSQGSFEYQDTLDHDYGGRASQPAADGTPAGLGATVYEEILQTSQSIARMRQASSRDLAFTEDKKKEKQFLNAESAYMDPMKQNGGPLTPGTSPTQLAAPVSFPTSTSSDSSGGRVIPDVRVTQHFAKEPQEPLKLHSSPASPSLASKEVGMTFSQGPGTPATTAMAPCPASLPRGYMTPAGPERSPSTSSTIHSYGQPPTTANYGSQTEELPHAPSGPAGSGRASREKPLSGGDGEVGPPQPSRGYSYFTGSSPPLSPSTPSESPTFSPSKLGPRATAEFSTQTPSLTPSSDIPRSVGTPSPMVAQGTQTPHRPSTPRLVWQQSSQEAPVMVITLASDASSQTRMVHASASTSPLCSPTDSQPASHSYSQTTPPSASQMPSEPAGPPGFPRAPSAGVDGPLALYGWGALPAENISLCRISSVPGTSRVEPGPRPPGTAVVDLRTAVKPTPIILTDQGMDLTSLAVEARKYGLALDPVPGRQSTAVQPLVINLNAQEQTHTFLATATTVSITMASSVLMAQQKQPVVYGDPFQSRLDFGQGSGSPVCLAQVKQVEQAVQTAPYRGGPRGRPREAKFARYNLPNQVTPLARRDILITQMGTAQSVSLKPGPVPEPGAEPHRATPAELRAHALPGTRKPHTVVVQMGEGAAGTVTTLLPEEPAGALDLTGMRPESRLACCDMAYKFPFGSSCTGTFHPAPSAPDKSVTDAALPGQSSGPFYSPRDPEPPEPLTFRAQGVVGPGPHEEQRPYPQGLPGRLYSSMSDTNLAEAGLNYHAQRIGQLFQGPGRDSAVDLSSLKHSYSLGFADGRYLGQGLQYGSFTDLRHPTDLLSHPLPMRPYSSVSNIYSDHRYGPRGDAVGFQEASLAQYSATTAREISRMCAALNSMDQYGGRHGGGSGGPDLVPYQPQHGPGLNAPQGLASLRSGLLGNPTYPEGQPSPGNLAQYGPAASQGTAVRQLLPSTATVRAADGMIYSTINTPIAATLPITTQPASVLRPMVRGGMYRPYGSGGVTAVPLTSLTRVPMIAPRVPLGPAGLYRYPAPSRFPIASTIPPAEGPVYLGKPAAAKASGAGGPPRPELPAGGAREEPLSTTAPPAVIKEAPVAQAPAPPPGQKPAGDAAAGSGSGVLGRPVMEKEEASQEDRQRKQQEQLLQLERERVELEKLRQLRLQEELERERVELQRHREEEQLLVQRELQELQTIKHHVLQQQQEERQAQFALQREQLAQQRLQLEQIQQLQQQLQQQLEEQKQRQKAPFPATCEAPSRGPPPAATELAQNGQYWPPLTHTAFIAVAGTEGPGQAREPVLHRGLPSSASDMSLQTEEQWEAGRSGIKKRHSMPRLRDACEPESGPDPSTVRRIADSSVQTDDEEGEGRYLLTRRRRTRRSADCSVQTDDEDNAEWEQPVRRRRSRLSRHSDSGSDSKHEASASSSAAAAAARAMSSVGIQTISDCSVQTEPEQLPRVSPAIHITAATDPKVEIVRYISAPEKTGRGESLACQTEPDGQAQGVAGPQLIGPTAISPYLPGIQIVTPGALGRFEKKKPDPLEIGYQAHLPPESLSQLVSRQPPKSPQVLYSPVSPLSPHRLLDTSFASSERLNKAHVSPQKQFIADSTLRQQTLPRPMKTLQRSLSDPKPLSPTAEESAKERFSLYQHQGGLGSQVSALPPNGLVRKVKRTLPSPPPEEAHLPLAGQVPSQLYAASLLQRGLAGPTTVPATKASLLRELDRDLRLVEHESTKLRKKQAELDEEEKEIDAKLKYLELGITQRKESLAKDRVGRDYPPLRGLGEHRDYLSDSELNQLRLQGCTTPAGQYVDYPASAAVPATPSGPTAFQQPRFPPAATQYTAGSSGPTQNGFLAHQAPTYTGPSTYPAPTYPPGTSYPAEPGLPSQPAFHPTGHYAAPTPMPTTQSAPFPVQADSHAAHQKPRQTSLADLEQKVPTNYEVISSPAVTVSSTPSETGYSGPAVSSSYEHGKAPEHPRGGDRSSVSQSPAPTYPSDSHYTSLEQNVPRNYVMIDDISELTKDSTPTASDSQRPEPLGPGGVSGRPGKDPGEPAVLEGPTLPCCYGRGEEESEEDSYDPRGKSGHHRSMESNGRPASTHYYSDSDYRHGARADKYGPGPMGPKHPSKNLAPAAISSKRSKHRKQGMEQKISKFSPIEEAKDVESDLASYPPPTVSSSLTSRSRKFQDEITYGLKKNVYEQQRYYGVSSRDTAEEDDRMYGGSSRSRVASAYSGEKLSSHDFSSRSKGYERERETAQRLQKAGPKPSSLSMAHGRARPPMRSQASEEESPVSPLGRPRPAGGALPPGDTCPQFCSSHSMPDVQEHVKDGPRAHAYKREEGYILDDSHCVVSDSEAYHLGQEETDWFDKPRDARSDRFRHHGGHTVSSSQKRGPARHSYHDYDEPPEEGLWPHDEGGPGRHTSAKEHRHHGDHGRHSGRHAGEEPGRRAARPHARDMGRHETRPHPQASPAPAMQKKGQPGYPSSADYSQPSRAPSAYHHASDSKKGSRQAHSGPTVLQPKPEAQAQPQMQGRQAVPGPQQSQPPSSRQTPSGTASRQPQTQQQQQQQQQQQQQQQQQQQQQQQQGLGQQAPQQAPSQARLQQQSQPTTRSTAPAASHPAGKPQPGPTTAPGPQPAGLPRAEQAGSSKPAAKAPQQGRAPQAQSAPGPAGAKTGARPGGTPGAPAGQPAAEGESVFSKILPGGAAEQAGKLTEAVSAFGKKFSSFW 5BF3C230E2C71AE2 pclo-rabac1 2 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait region zn2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association we employed Pic-Zn2 as a bait in a yeast two-hybrid screen to identify potential interacting partners. Screening 3 106 transformants yielded seven independent clones interacting with Pic-Zn2. Two clones encoded PRA1, a small 21 kDa protein originally identified through its interaction with rab3A pclo-rabac1-1 3 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait region zn2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 5A As shown in Figure 5A, PRA1 could interact with Pic-Zn2 but not Rabp-Zn. rab3a-rabp 3 12 unspecified role unspecified role bait bait n terminal optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain 15 unspecified role unspecified role prey prey physical association physical association 5A Only yeast doubly transfected with rab3A and Rabp-Zn were able to grow on plates lacking histidine and turned blue in 30 min when incubated with X-gal ( Figure 5A). pclo-rabac1-2 5 8 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 9 unspecified role unspecified role prey prey zn1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain zn2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 17 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction 5B As seen in Figure 5B, His6-PRA1 interacted with both GST-Pic-Zn1 and GST-Pic-Zn2, but not with GST-Rabp-Zn or GST. pclo-rabac1-3 4 8 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait zn finger 2 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain zn finger 1 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction 5D ELISA binding assay demonstrating the ability of PRA1 to interact specifically with the Piccolo zinc fingers. Triton X-100 pic-bsn 1 18 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component colocalization colocalization 4D, 4E, 4F Arrows point to examples of colocalizing clusters. Approximately 95% and 99% of the Piccolo (Pic) clusters (A and D) are observed to colocalize with synaptotagmin (Stg) (C) and Bassoon (Bas) (E) clusters along dendritic profiles, respectively. Piccolo clusters (G) are also observed to colocalize with GluR1 (GluR) subunits of the AMPA receptor at axodendritic and axospinous synapses. Piccolo (J) clusters are also colocalized with GABAA Receptor (GABAR) (K) clusters at axodendritic and axosomatic synapses. pic-glur 1 16 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component colocalization colocalization 4G, 4H, 4I Arrows point to examples of colocalizing clusters. Approximately 95% and 99% of the Piccolo (Pic) clusters (A and D) are observed to colocalize with synaptotagmin (Stg) (C) and Bassoon (Bas) (E) clusters along dendritic profiles, respectively. Piccolo clusters (G) are also observed to colocalize with GluR1 (GluR) subunits of the AMPA receptor at axodendritic and axospinous synapses. Piccolo (J) clusters are also colocalized with GABAA Receptor (GABAR) (K) clusters at axodendritic and axosomatic synapses. pic-pra1 1 10 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component colocalization colocalization 6A PRA1 clusters are found colocalized with Piccolo and synaptotagmin clusters along dendritic profiles pra1-syp 6 14 unspecified role unspecified role neutral component neutral component 10 unspecified role unspecified role neutral component neutral component association association 6C To assess whether PRA1 is associated with SVs, we performed a flotation assay with lysed synaptosomes. Similar to synaptophysin, PRA1 immunoreactivity was found in the 0.3 M sucrose fraction, indicating that PRA1 is associated with SVs ( Figure 6C). pra1-syn 7 11 unspecified role unspecified role prey prey 10 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey 14 unspecified role unspecified role bait bait acetylation acetylation reaction acetylation 6D In the bound fraction, PRA1 was affinity purified along with synaptophysin, rab3A and VAMP/Synaptobrevin II ( Figure 6D).