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/ Isolation and characterization of a dual prenylated Rab and VAMP2 receptor. Isolation and characterization of a dual prenylated Rab and VAMP2 receptor. J. Biol. Chem. (0021-9258) 1997 Martincic I., Peralta ME., Ngsee JK. Synapse - Interactions of proteins with an established role in the presynapse. Source of Rab3A inferred from other work by the same author ngsee@civich.ottawa.on.ca 09-April-2009: Contacted by JYOTI. rat brain cDNA library orchard in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Martincic I., Peralta ME., Ngsee JK. Synapse - Interactions of proteins with an established role in the presynapse. Source of Rab3A inferred from other work by the same author ngsee@civich.ottawa.on.ca Accepted 2009-MAR-26 by ORCHARD. 09-April-2009: Contacted by JYOTI. rat brain cDNA library J. Biol. Chem. (0021-9258) 1997 orchard Isolation and characterization of a dual prenylated Rab and VAMP2 receptor. Isolation and characterization of a dual prenylated Rab and VAMP2 receptor. J. Biol. Chem. (0021-9258) 1997 Martincic I., Peralta ME., Ngsee JK. Synapse - Interactions of proteins with an established role in the presynapse. Source of Rab3A inferred from other work by the same author ngsee@civich.ottawa.on.ca 09-April-2009: Contacted by JYOTI. rat brain cDNA library orchard 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 Martincic I., Peralta ME., Ngsee JK. Synapse - Interactions of proteins with an established role in the presynapse. Source of Rab3A inferred from other work by the same author ngsee@civich.ottawa.on.ca Accepted 2009-MAR-26 by ORCHARD. 09-April-2009: Contacted by JYOTI. rat brain cDNA library J. Biol. Chem. (0021-9258) 1997 orchard Isolation and characterization of a dual prenylated Rab and VAMP2 receptor. Isolation and characterization of a dual prenylated Rab and VAMP2 receptor. J. Biol. Chem. (0021-9258) 1997 Martincic I., Peralta ME., Ngsee JK. Synapse - Interactions of proteins with an established role in the presynapse. Source of Rab3A inferred from other work by the same author ngsee@civich.ottawa.on.ca 09-April-2009: Contacted by JYOTI. rat brain cDNA library orchard 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 Martincic I., Peralta ME., Ngsee JK. Synapse - Interactions of proteins with an established role in the presynapse. The organism of origin of Vamp proteins cannot be deciphered from the publication, however, the rat and human proteins are very similar hence the organism of the protein assumed to be rat. no when peptide origin unknown assumed to be that of the rest of the interacting proteins. I am hoping someone concerned with the entry does come back with the organism in due course. Source of Rab3A inferred from other work by the same author ngsee@civich.ottawa.on.ca Accepted 2009-MAR-26 by ORCHARD. 09-April-2009: Contacted by JYOTI. rat brain cDNA library J. Biol. Chem. (0021-9258) 1997 orchard stx1a_rat Syntaxin-1A Synaptotagmin-associated 35 kDa protein Neuron-specific antigen HPC-1 Stx1a Sap protein protein rat Rattus norvegicus (Rat) Rat MKDRTQELRTAKDSDDDDDVTVTVDRDRFMDEFFEQVEEIRGFIDKIAENVEEVKRKHSAILASPNPDEKTKEELEELMSDIKKTANKVRSKLKSIEQSIEQEEGLNRSSADLRIRKTQHSTLSRKFVEVMSEYNATQSDYRERCKGRIQRQLEITGRTTTSEELEDMLESGNPAIFASGIIMDSSISKQALSEIETRHSEIIKLENSIRELHDMFMDMAMLVESQGEMIDRIEYNVEHAVDYVERAVSDTKKAVKYQSKARRKKIMIIICCVILGIIIASTIGGIFG DIP protein P32851 original sequence version: 74 DIP protein P32851 original sequence version: 78 DIP protein P32851 original sequence version: 142 DIP protein P32851 original sequence version: 138 DIP protein P32851 original sequence version: 89 DIP protein P32851 original sequence version: 115 DIP protein P32851 original sequence version: 134 DIP protein P32851 original sequence version: 121 DIP protein P32851 original sequence version: 123 DIP protein P32851 original sequence version: 150 DIP protein P32851 original sequence version: 156 DIP protein P32851 original sequence version: 163 DIP protein P32851 original sequence version: 126 DIP protein P32851 original sequence version: 124 DIP protein P32851 original sequence version: 158 8F929F489E323CAA vamp1_rat Vesicle-associated membrane protein 1 Vamp1 Syb1 Synaptobrevin-1 protein protein rat Rattus norvegicus (Rat) Rat MSAPAQPPAEGTEGAAPGGGPPGPPPNTTSNRRLQQTQAQVEEVVDIMRVNVDKVLERDQKLSELDDRADALQAGASVFESSAAKLKRKYWWKNCKMMIMLGAICAIIVVVIVIYIFT E34D4D735CFBF6A8 praf1_rat Prenylated Rab acceptor protein 1 Rabac1 Pra1 Praf1 PRA1 family protein 1 protein protein rat Rattus norvegicus (Rat) Rat MAAQKDQQKDAEVEGLSATTLLPKLIPSGAGREWLERRRATIRPWGTFVDQQRFSRPRNVGELCQRLVRNVEYYQSNYVFVFLGLILYCVVTSPMLLVALAVFFGACYILYLRTLQSKLVLFGREVSPAHQYALAGGVSFPFFWLAGAGSAVFWVLGATLVLIGSHAAFHQIEPADGEELQMEPV DA6341AE66F5C2F0 rab1a_rat Ras-related protein Rab-1A Rab1A Rab1 protein protein rat Rattus norvegicus (Rat) Rat MSSMNPEYDYLFKLLLIGDSGVGKSCLLLRFADDTYTESYISTIGVDFKIRTIELDGKTIKLQIWDTAGQERFRTITSSYYRGAHGIIVVYDVTDQESFNNVKQWLQEIDRYASENVNKLLVGNKCDLTTKKVVDYTTAKEFADSLGIPFLETSAKNATNVEQSFMTMAAEIKKRMGPGATAGGAEKSNVKIQSTPVKQSGGGCC B2A8F4E3B0FB17D6 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 vamp2-vamp1_protein fusion protein between Vamp2 1-31 and Vamp1 34-118 protein protein rat Rattus norvegicus (Rat) Rat MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVEEVVDIMRVNVDKVLERDQKLSELDDRADALQAGASVFESSAAKLKRKYWWKNCKMMIMLGAICAIIVVVIVIYIFT An amino acid sequence which is not represented in UniProt 911C2AC066821945 rab3i_rat Rab-3A-interacting protein Rab3ip Rabin3 Rabin-3 SSX2-interacting protein Rabin8 protein protein rat Rattus norvegicus (Rat) Rat MANDPLEGFHEVNLASPTSPDLLGVCDPGTQEQTTSPSVIYRPHPSTLCSATIQANALNLSDLPTQPVYSSPRHLNCAEISNISIHVPEPASSVASEVAAGLTRFTSRKDSCNAEREFLQGATVTEASAGNDDIFGLSTDSLSRLRSPSVLEVREKGYERLKEELAKAQRELKLKDEECERLSKVRDQLGQELEELTASLFEEAHKMVREANVKQATAEKQLKEAQGKIDVLQAEVAALKTLVLSSSPTSPTQEPLAAGKTPFKRGHTRNKSTSSAMSGSHQDFSAIQPIVKDCREADLSLYNEFRSWKDEPTMDRTCPFLDKIYQEDIFPCLTFAKSELASAVLEAVENNTLSIEPVGLQPIRFVKASAVECGGPKKCALTGQSKPCKHRIKLGDSSSYYYISPVCRYRITSVCNFFTYIRYIQQGLVKQQDVDQMFWEVMQLRKEMSLAKLGYFKEEL 6DFAA787292E7577 rab3a_rat Ras-related protein Rab-3A Rab3a protein protein rat Rattus norvegicus (Rat) Rat MASATDSRYGQKESSDQNFDYMFKILIIGNSSVGKTSFLFRYADDSFTPAFVSTVGIDFKVKTIYRNDKRIKLQIWDTAGQERYRTITTAYYRGAMGFILMYDITNEESFNAVQDWSTQIKTYSWDNAQVLLVGNKCDMEDERVVSSERGRQLADHLGFEFFEASAKDNINVKQTFERLVDVICEKMSESLDTADPAVTGAKQGPQLTDQQAPPHQDCAC 1A3E9F8C9D09EB40 rabac1-rab3a 2 11 unspecified role unspecified role bait bait P63012:p.Gln81Leu mutation mutation certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 19:03:18 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln81leu' Tue Jun 30 19:45:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln81Leu' 6 unspecified role unspecified role prey prey physical association physical association Our initial analysis from the yeast two-hybrid screen indicates that PRA1 interacts with both Rab3A and Rab1. rabac1-rab1 3 6 unspecified role unspecified role unspecified role unspecified role 7 unspecified role unspecified role unspecified role unspecified role Q6NYB7:p.Cys204_Cys205del mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain CC .. Wed Jul 06 16:28:29 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'c terminal' Wed May 02 18:48:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys_cys204-205del_del' Tue Jun 30 16:06:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys204_Cys205del' physical association physical association Our initial analysis from the yeast two-hybrid screen indicates that PRA1 interacts with both Rab3A and Rab1. 4 rabac1-rab3a-1 3 11 unspecified role unspecified role bait bait P63012:p.Cys218_Cys220del mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain CAC ... Wed Jul 06 16:27:09 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'region' Wed May 02 19:48:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys_ala_cys218-220del_del_del' Tue Jun 30 17:43:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys218_Cys220del' 6 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 4, 5, 6 When the carboxyl-terminal CXC of Rab3A was mutated to CVLL and CVLS interaction was observed with Praf1. This indicates that PRA1 is capable of recognizing Rab with a mono-GG group and that interaction is not limited to GG-modified Rab but also includes farnesylated Rab. Deletion of the carboxyl-terminal CXC prenylation signal in Rab3A (Rab3A(Delta CXC), and Rab3A(Q81L,Delta CXC)), and the last two cysteines in Rab1 (Rab1(Delta CC)) completely abrogated PRA1 interaction (Fig. 4). rain_3-rab3a 2 11 unspecified role unspecified role bait bait P63012:p.Gln81Leu mutation mutation certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 17:07:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln81leu' Tue Jun 30 17:52:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln81Leu' 10 unspecified role unspecified role prey prey physical association physical association rabac1-vamp2 3 8 unspecified role unspecified role unspecified role unspecified role tmd required to bind necessary binding region required to bind range range range range Wed Jul 06 16:30:17 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'tmd' 6 unspecified role unspecified role unspecified role unspecified role region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. 5A shows that PRA1 interacted with VAMP2 but not with closely related VAMP1 or the ubiquitous cellubrevin. 5, 6 rabac1-vamp2_vamp1 3 9 unspecified role unspecified role unspecified role unspecified role 6 unspecified role unspecified role unspecified role unspecified role physical association physical association 5 This VAMP2-VAMP1 chimera showed a positive beta -galactosidase reaction when tested against PRA1. syntaxin1a-vamp2 3 4 unspecified role unspecified role unspecified role unspecified role 8 unspecified role unspecified role unspecified role unspecified role physical association physical association Both VAMP1 and VAMP2 interacted strongly with Syn1A, whereas a weaker interaction was detected with cellubrevin (Fig. 5B). 5 syntaxin1a-vamp1 3 5 unspecified role unspecified role unspecified role unspecified role 4 unspecified role unspecified role unspecified role unspecified role physical association physical association Both VAMP1 and VAMP2 interacted strongly with Syn1A, whereas a weaker interaction was detected with cellubrevin (Fig. 5B). 5 rabac1-vamp2-1 1 6 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range 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 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role bait bait tmd required to bind necessary binding region required to bind range range range range Wed Jul 06 16:14:22 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'tmd' 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 7,8 PRA1(1-185) can bind to the immobilized full-length VAMP2 We estimated a 20-fold difference in the binding of the full-length PRA1(1-185) to immobilized GST-VAMP2 when compared with the truncated PRA1(1-164) (Fig. 8B). The C-terminally truncated form of Rabac1 bound 20 times lesser to Vamp2 Coordinates of the transmembrane domain has not been given in the text, but instead inferred from the UniProtKB entry