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/ Identification and characterization of human VCY2-interacting protein: VCY2IP-1, a microtubule-associated protein-like protein. Identification and characterization of human VCY2-interacting protein: VCY2IP-1, a microtubule-associated protein-like protein. Biol. Reprod. (0006-3363) 2004 imex curation Wong EY., Tse JY., Yao KM., Lui VC., Tam PC., Yeung WS. Apoptosis - Interactions involving proteins with a function related to apoptosis jymtse@yahoo.com.hk 03-Feb-2009: Contacted by JYOTI. Human testis cDNA davet Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Wong EY., Tse JY., Yao KM., Lui VC., Tam PC., Yeung WS. Apoptosis - Interactions involving proteins with a function related to apoptosis jymtse@yahoo.com.hk Accepted 2009-FEB-13 by DAVET. 03-Feb-2009: Contacted by JYOTI. Human testis cDNA Biol. Reprod. (0006-3363) 2004 davet Only protein-protein interactions imex curation Identification and characterization of human VCY2-interacting protein: VCY2IP-1, a microtubule-associated protein-like protein. Identification and characterization of human VCY2-interacting protein: VCY2IP-1, a microtubule-associated protein-like protein. Biol. Reprod. (0006-3363) 2004 imex curation Wong EY., Tse JY., Yao KM., Lui VC., Tam PC., Yeung WS. Apoptosis - Interactions involving proteins with a function related to apoptosis jymtse@yahoo.com.hk 03-Feb-2009: Contacted by JYOTI. Human testis cDNA davet 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 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Wong EY., Tse JY., Yao KM., Lui VC., Tam PC., Yeung WS. Apoptosis - Interactions involving proteins with a function related to apoptosis jymtse@yahoo.com.hk Accepted 2009-FEB-13 by DAVET. 03-Mar-2009: Contacted by JYOTI. Human testis cDNA Biol. Reprod. (0006-3363) 2004 davet Only protein-protein interactions imex curation Identification and characterization of human VCY2-interacting protein: VCY2IP-1, a microtubule-associated protein-like protein. Identification and characterization of human VCY2-interacting protein: VCY2IP-1, a microtubule-associated protein-like protein. Biol. Reprod. (0006-3363) 2004 imex curation Wong EY., Tse JY., Yao KM., Lui VC., Tam PC., Yeung WS. Apoptosis - Interactions involving proteins with a function related to apoptosis jymtse@yahoo.com.hk 03-Feb-2009: Contacted by JYOTI. Human testis cDNA davet Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast two hybrid pooling two hybrid pooling approach two hybrid pooling predetermined predetermined participant predetermined Wong EY., Tse JY., Yao KM., Lui VC., Tam PC., Yeung WS. Apoptosis - Interactions involving proteins with a function related to apoptosis jymtse@yahoo.com.hk Accepted 2009-FEB-13 by DAVET. 03-Feb-2009: Contacted by JYOTI. Human testis cDNA Biol. Reprod. (0006-3363) 2004 davet Only protein-protein interactions imex curation vcy2_human Testis-specific basic protein Y 2 BPY2C BPY2A BPY2 BPY2B VCY2 VCY2A VCY2B VCY2C Variably charged protein Y 2 Basic charge, Y-linked 2 protein protein human Homo sapiens Human MMTLVPRARTRAGQDHYSHPCPRFSQVLLTEGIMTYCLTKNLSDVNILHRLLKNGNVRNTLLQSKVGLLTYYVKLYPGEVTLLTRPSIQMRLCCITGSVSRPRSQK 2573EC02A2C99788 map1s_human Microtubule-associated protein 1S MAP1S BPY2IP1 C19orf5 MAP8 VCY2IP1 BPY2-interacting protein 1 Microtubule-associated protein 8 Variable charge Y chromosome 2-interacting protein 1 protein protein human Homo sapiens Human MAAVAGSGAAAAPSSLLLVVGSEFGSPGLLTYVLEELERGIRSWDVDPGVCNLDEQLKVFVSRHSATFSSIVKGQRSLHHRGDNLETLVLLNPSDKSLYDELRNLLLDPASHKLLVLAGPCLEETGELLLQTGGFSPHHFLQVLKDREIRDILATTPPPVQPPILTITCPTFGDWAQLAPAVPGLQGALRLQLRLNPPAQLPNSEGLCEFLEYVAESLEPPSPFELLEPPTSGGFLRLGRPCCYIFPGGLGDAAFFAVNGFTVLVNGGSNPKSSFWKLVRHLDRVDAVLVTHPGADSLPGLNSLLRRKLAERSEVAAGGGSWDDRLRRLISPNLGVVFFNACEAASRLARGEDEAELALSLLAQLGITPLPLSRGPVPAKPTVLFEKMGVGRLDMYVLHPPSAGAERTLASVCALLVWHPAGPGEKVVRVLFPGCTPPACLLDGLVRLQHLRFLREPVVTPQDLEGPGRAESKESVGSRDSSKREGLLATHPRPGQERPGVARKEPARAEAPRKTEKEAKTPRELKKDPKPSVSRTQPREVRRAASSVPNLKKTNAQAAPKPRKAPSTSHSGFPPVANGPRSPPSLRCGEASPPSAACGSPASQLVATPSLELGPIPAGEEKALELPLAASSIPRPRTPSPESHRSPAEGSERLSLSPLRGGEAGPDASPTVTTPTVTTPSLPAEVGSPHSTEVDESLSVSFEQVLPPSAPTSEAGLSLPLRGPRARRSASPHDVDLCLVSPCEFEHRKAVPMAPAPASPGSSNDSSARSQERAGGLGAEETPPTSVSESLPTLSDSDPVPLAPGAADSDEDTEGFGVPRHDPLPDPLKVPPPLPDPSSICMVDPEMLPPKTARQTENVSRTRKPLARPNSRAAAPKATPVAAAKTKGLAGGDRASRPLSARSEPSEKGGRAPLSRKSSTPKTATRGPSGSASSRPGVSATPPKSPVYLDLAYLPSGSSAHLVDEEFFQRVRALCYVISGQDQRKEEGMRAVLDALLASKQHWDRDLQVTLIPTFDSVAMHTWYAETHARHQALGITVLGSNSMVSMQDDAFPACKVEF 30AB33FFE26DDF91 by2-map1s-1 2 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association A bait vector encoding full-length VCY2 was used in yeast two-hybrid screens of a human testis cDNA library. Twenty-six positive-interacting clones were isolated and characterized, 16 of which encoded the same 948-bp cDNA insert. This VCY2-interacting clone (MHC1) was named VCY2IP-1 (VCY2-interacting protein-1). by2-map1s-2 3 4 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association The specificity of the interaction between VCY2IP-1 and VCY2 was further confirmed by testing interactions between specific proteins during yeast mating using two different yeast strains. Fig. 1 by2-map1s-3 1 4 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED 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 in vitro In vitro 5 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 region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro direct interaction direct interaction Coimmunoprecipitation of VCY2 and VCY2IP-1 in vitro-transcribed/translated proteins Fig. 2