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/ Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Oncogene (0950-9232) 2003 Ohno H., Hirabayashi S., Kansaku A., Yao I., Tajima M., Nishimura W., Ohnishi H., Mashima H., Fujita T., Omata M., Hata Y. Synapse - Interactions of proteins with an established role in the presynapse. yuhammch@med.tmd.ac.jp 26-APR-2007: Contacted by JYOTI. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Ohno H., Hirabayashi S., Kansaku A., Yao I., Tajima M., Nishimura W., Ohnishi H., Mashima H., Fujita T., Omata M., Hata Y. Synapse - Interactions of proteins with an established role in the presynapse. yuhammch@med.tmd.ac.jp Accepted 2007-MAY-10 by JYOTI. 26-APR-2007: Contacted by JYOTI. Oncogene (0950-9232) 2003 Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Oncogene (0950-9232) 2003 Ohno H., Hirabayashi S., Kansaku A., Yao I., Tajima M., Nishimura W., Ohnishi H., Mashima H., Fujita T., Omata M., Hata Y. Synapse - Interactions of proteins with an established role in the presynapse. yuhammch@med.tmd.ac.jp 26-APR-2007: Contacted by JYOTI. canlf-mdck Canis familiaris (dog) kidney epithelial cells Canis lupus familiaris Dog Canis familiaris mdck Canine Cocker Spaniel kidney epithelial cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ohno H., Hirabayashi S., Kansaku A., Yao I., Tajima M., Nishimura W., Ohnishi H., Mashima H., Fujita T., Omata M., Hata Y. Synapse - Interactions of proteins with an established role in the presynapse. Semiconfluent MDCK cells were extracted using urea yuhammch@med.tmd.ac.jp Accepted 2007-MAY-10 by JYOTI. 26-APR-2007: Contacted by JYOTI. Oncogene (0950-9232) 2003 Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Oncogene (0950-9232) 2003 Ohno H., Hirabayashi S., Kansaku A., Yao I., Tajima M., Nishimura W., Ohnishi H., Mashima H., Fujita T., Omata M., Hata Y. Synapse - Interactions of proteins with an established role in the presynapse. yuhammch@med.tmd.ac.jp 26-APR-2007: Contacted by JYOTI. canlf-mdck Canis familiaris (dog) kidney epithelial cells Canis lupus familiaris Dog Canis familiaris mdck Canine Cocker Spaniel kidney epithelial cell line confocal microscopy confocal microscopy immunostaining immunostaining Ohno H., Hirabayashi S., Kansaku A., Yao I., Tajima M., Nishimura W., Ohnishi H., Mashima H., Fujita T., Omata M., Hata Y. Synapse - Interactions of proteins with an established role in the presynapse. Cells treated with EGTA to encourage separation and then in the presence of calcium at which point immature cell contacts begin to form. yuhammch@med.tmd.ac.jp Accepted 2007-MAY-10 by JYOTI. 26-APR-2007: Contacted by JYOTI. Oncogene (0950-9232) 2003 Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Oncogene (0950-9232) 2003 Ohno H., Hirabayashi S., Kansaku A., Yao I., Tajima M., Nishimura W., Ohnishi H., Mashima H., Fujita T., Omata M., Hata Y. Synapse - Interactions of proteins with an established role in the presynapse. yuhammch@med.tmd.ac.jp 26-APR-2007: Contacted by JYOTI. 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 Ohno H., Hirabayashi S., Kansaku A., Yao I., Tajima M., Nishimura W., Ohnishi H., Mashima H., Fujita T., Omata M., Hata Y. Synapse - Interactions of proteins with an established role in the presynapse. yuhammch@med.tmd.ac.jp Accepted 2007-MAY-10 by JYOTI. 26-APR-2007: Contacted by JYOTI. Oncogene (0950-9232) 2003 Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Carom: a novel membrane-associated guanylate kinase-interacting protein with two SH3 domains. Oncogene (0950-9232) 2003 Ohno H., Hirabayashi S., Kansaku A., Yao I., Tajima M., Nishimura W., Ohnishi H., Mashima H., Fujita T., Omata M., Hata Y. Synapse - Interactions of proteins with an established role in the presynapse. yuhammch@med.tmd.ac.jp 26-APR-2007: Contacted by JYOTI. chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Ohno H., Hirabayashi S., Kansaku A., Yao I., Tajima M., Nishimura W., Ohnishi H., Mashima H., Fujita T., Omata M., Hata Y. Synapse - Interactions of proteins with an established role in the presynapse. yuhammch@med.tmd.ac.jp Accepted 2007-MAY-10 by JYOTI. 26-APR-2007: Contacted by JYOTI. Oncogene (0950-9232) 2003 cask_canfa_protein Canis familiaris CASK protein protein canlf Canis familiaris (dog) Canis lupus familiaris Dog Canis familiaris An amino acid sequence which is not represented in UniProt Similar to P07498 fcsd2_human F-BAR and double SH3 domains protein 2 FCHSD2 KIAA0769 SH3MD3 Protein nervous wreck 1 SH3 multiple domains protein 3 Carom protein protein human Homo sapiens Human MQPPPRKVKVTQELKNIQVEQMTKLQAKHQAECDLLEDMRTFSQKKAAIEREYAQGMQKLASQYLKRDWPGVKADDRNDYRSMYPVWKSFLEGTMQVAQSRMNICENYKNFISEPARTVRSLKEQQLKRCVDQLTKIQTELQETVKDLAKGKKKYFETEQMAHAVREKADIEAKSKLSLFQSRISLQKASVKLKARRSECNSKATHARNDYLLTLAAANAHQDRYYQTDLVNIMKALDGNVYDHLKDYLIAFSRTELETCQAVQNTFQFLLENSSKVVRDYNLQLFLQENAVFHKPQPFQFQPCDSDTSRQLESETGTTEEHSLNKEARKWATRVAREHKNIVHQQRVLNDLECHGAAVSEQSRAELEQKIDEARENIRKAEIIKLKAEARLDLLKQIGVSVDTWLKSAMNQVMEELENERWARPPAVTSNGTLHSLNADTEREEGEEFEDNMDVFDDSSSSPSGTLRNYPLTCKVVYSYKASQPDELTIEEHEVLEVIEDGDMEDWVKARNKVGQVGYVPEKYLQFPTSNSLLSMLQSLAALDSRSHTSSNSTEAELVSGSLNGDASVCFVKALYDYEGQTDDELSFPEGAIIRILNKENQDDDGFWEGEFNGRIGVFPSVLVEELSASENGDTPWMREIQISPSPKPHASLPPLPLYDQPPSSPYPSPDKRSSLYFPRSPSANEKSLHAESPGFSQASRHTPETSYGKLRPVRAAPPPPTQNHRRPAEKIEDVEITLV 3AF4DAC3CAD24C28 magi1_canfa_protein Canis familiaris membrane-associated guanylate kinase, WW and PDZ domain-containing protein 1 protein protein canlf Canis familiaris (dog) Canis lupus familiaris Dog Canis familiaris An amino acid sequence which is not represented in UniProt Similar to Q96QZ7 cskp_rat Peripheral plasma membrane protein CASK Calcium/calmodulin-dependent serine protein kinase Cask protein protein rat Rattus norvegicus (Rat) Rat MADDDVLFEDVYELCEVIGKGPFSVVRRCINRETGQQFAVKIVDVAKFTSSPGLSTEDLKREASICHMLKHPHIVELLETYSSDGMLYMVFEFMDGADLCFEIVKRADAGFVYSEAVASHYMRQILEALRYCHDNNIIHRDVKPHCVLLASKENSAPVKLGGFGVAIQLGESGLVAGGRVGTPHFMAPEVVKREPYGKPVDVWGCGVILFILLSGCLPFYGTKERLFEGIIKGKYKMNPRQWSHISESAKDLVRRMLMLDPAERITVYEALNHPWLKERDRYAYKIHLPETVEQLRKFNARRKLKGAVLAAVSSHKFNSFYGDPPEELPDFSEDPTSSGLLAAERAVSQVLDSLEEIHALTDCSEKDLDFLHSVFQDQHLHTLLDLYDKINTKSSPQIRNPPSDAVQRAKEVLEEISCYPENNDAKELKRILTQPHFMALLQTHHVVAHEVYSDEALRVTPPPTSPYLNGDSPESANGDMDMENVTRVRLVQFQKNTDEPMGITLKMNELNHCIVARIMHGGMIHRQGTLHVGDEIREINGISVANQTVEQLQKMLREMRGSITFKIVPSYRTQSSSCERDSPSTSRQSPANGHSSTNNSVSIYVRAQFEYDPAKDDLIPCKEAGIRFRVGDIIQIISKDDHNWWQGKLENSKNGTAGLIPSPELQEWRVACIAMEKTKQEQQASCTWFGKKKKQYKDKYLAKHNADLVTYEEVVKLPAFKRKTLVLLGAHGVGRRHIKNTLITKHPERFAYPIPHTTRPPKKDEENGKNYYFVSHDQMMQDISNNEYLEYGSHEDAMYGTKLETIRKIHEQGLIAILDVEPQALKVLRTAEFPPFVVFIAAPTITPGLNEDESLQRLQKESDVLQRTYAHYFDLTIINNEIDETIRHLEEAVELVCTAPQWVPVSWVY 3694A9A44AC29629 f1ptw8_canlf FCHSD2 protein protein canlf Canis familiaris (dog) Canis lupus familiaris Dog Canis familiaris MQPPPRKVKVTQELKNTQVEQMTKLQAKHQAECDLLEDMRTFSQKKAAIEREYAQGIQKLASQYLKRDWPGIKADDRIDYRSMYPVWKSFLEGTMQVAQSRINICENYKNFISEPARTVRSIKEQQLKRCVDQLTKIQTELQETVKDLAKGKKKYFETEQMAHAVREKADIEAKSKLSLFQSRISLQKASVKLKARRSECNSKATHARNDYLLTLAAANAHQDRYYQTDLVNIMKALDGNVYDHLKDYLIAFSRTELETCQAVQNTFQFLLETSSKVVRDYNLQLFLQENAVFHKPQPFQFQPCDSDTSLKAAQLVDIELSPVSALRMTIAESRQLESETGTTEEHSLNKEARKWATRVAREHKNIVHQQRVLDDLECHGVAVSEQSRAELEQKIDEARENIRKAEIIKLKAEARLDLLKQIGVSVDTWLKSAMNQVMEELENERWARLPAVTNNDTLHSLNADPEREEGEEFEDNMDVFDDSSSSPSGTLRNYPLTCKVVYSYKASQPDELTIEEHEVLEVIEDGDMEDWVKARNKVGQVGYVPEKYLQFPTSNSLLSMLQSLAALDSRSHTSSNSTEAELVSGSLNGDASVCFVKALYDYEGQTDDELSFPEGAIIRILNKENQDDDGFWEGEFSGRIGVFPSVLVEELSASENGDTPWMREIQISPSSKPPGSLPPLPLYDQPPSSPYPSPDKRSSQYFPRSPSANENSFLAESPGFSQAPRHTPETSYGKLRPVRAAPPPPTQNHRRPAEKIEDVEITLV E05F955B7CAD31EF magi1_human Membrane-associated guanylate kinase, WW and PDZ domain-containing protein 1 MAGI1 BAIAP1 BAP1 TNRC19 BAI1-associated protein 1 Membrane-associated guanylate kinase inverted 1 Atrophin-1-interacting protein 3 WW domain-containing protein 3 Trinucleotide repeat-containing gene 19 protein AIP3 protein protein human Homo sapiens Human MSKVIQKKNHWTSRVHECTVKRGPQGELGVTVLGGAEHGEFPYVGAVAAVEAAGLPGGGEGPRLGEGELLLEVQGVRVSGLPRYDVLGVIDSCKEAVTFKAVRQGGRLNKDLRHFLNQRFQKGSPDHELQQTIRDNLYRHAVPCTTRSPREGEVPGVDYNFLTVKEFLDLEQSGTLLEVGTYEGNYYGTPKPPSQPVSGKVITTDALHSLQSGSKQSTPKRTKSYNDMQNAGIVHAENEEEDDVPEMNSSFTADSGEQEEHTLQETALPPVNSSIIAAPITDPSQKFPQYLPLSAEDNLGPLPENWEMAYTENGEVYFIDHNTKTTSWLDPRCLNKQQKPLEECEDDEGVHTEELDSELELPAGWEKIEDPVYGIYYVDHINRKTQYENPVLEAKRKKQLEQQQQQQQQQQQQQQQQQQQQTEEWTEDHSALVPPVIPNHPPSNPEPAREVPLQGKPFFTRNPSELKGKFIHTKLRKSSRGFGFTVVGGDEPDEFLQIKSLVLDGPAALDGKMETGDVIVSVNDTCVLGHTHAQVVKIFQSIPIGASVDLELCRGYPLPFDPDDPNTSLVTSVAILDKEPIIVNGQETYDSPASHSSKTGKVNGMKDARPSSPADVASNSSHGYPNDTVSLASSIATQPELITVHIVKGPMGFGFTIADSPGGGGQRVKQIVDSPRCRGLKEGDLIVEVNKKNVQALTHNQVVDMLVECPKGSEVTLLVQRGGLPVPKKSPKSQPLERKDSQNSSQHSVSSHRSLHTASPSHSTQVLPEFPPAEAQAPDQTDSSGQKKPDPFKIWAQSRSMYENRPMSPSPASGLSKGEREREINSTNFGECPIPDYQEQDIFLWRKETGFGFRILGGNEPGEPIYIGHIVPLGAADTDGRLRSGDELICVDGTPVIGKSHQLVVQLMQQAAKQGHVNLTVRRKVVFAVPKTENEVPSPASSHHSSNQPASLTEEKRTPQGSQNSLNTVSSGSGSTSGIGSGGGGGSGVVSTVVQPYDVEIRRGENEGFGFVIVSSVSRPEAGTTFAGNACVAMPHKIGRIIEGSPADRCGKLKVGDRILAVNGCSITNKSHSDIVNLIKEAGNTVTLRIIPGDESSNATLLTNAEKIATITTTHTPSQQGTQETRNTTKPKQESQFEFKAPQATQEQDFYTVELERGAKGFGFSLRGGREYNMDLYVLRLAEDGPAERCGKMRIGDEILEINGETTKNMKHSRAIELIKNGGRRVRLFLKRGDGSVPEYDPSSDRHGPATGPQGVPEVRAGPDRRQHPSLESSYPPDLHKSSPHGEKRAHARDPKGSREYSRQPNEHHTWNGTSRKPDSGACRPKDRAPEGRRDAQAERAAAANGPKRRSPEKRREGTRSADNTLERREKHEKRRDVSPERRRERSPTRRRDGSPSRRRRSLERLLEQRRSPERRRGGSPERRAKSTDRRRARSPERRRERSLDKRNREDRASHREREEANLKQDAGRSSRHPPEQRRRPYKECSTDLSI 4B2EE05243A6FA65 magi1-fchsd2-1 4 7 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association We obtained the KIAA0769 gene product through the yeast two-hybrid screening using MAGI-1 as a bait and named it Carom. magi1-fchsd2-2 2 8 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association Coimmunoprecipitation of Carom with MAGI-1 from MDCK cells. 3A magi1-fchsd2-3 5 11 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 7 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 c-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association HA-MAGI-1 was coimmunoprecipitated with Myc-Carom-1 and -6 (Figure 3Ab, arrows) 3Ab From sequence length, FCHSD2 must be isoform -2 or -3 but not enough information given to decide which. Numbering recalculated to O94868 magi1-fchsd2-4 1 7 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 pdz binding region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 11 unspecified role unspecified role prey prey pdz region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 27 29 physical association physical association GST-Carom-1 captured Myc-MAGI-1-full and Myc-MAGI-V containing PDZ4 and PDZ5 (Figure 3Bb). GST-Carom-10 lacking the C-terminal PDZ-binding motif did not bind Myc-MAGI-1-V. 3Ba From sequence length, FCHSD2 must be isoform -2 or -3 but not enough information given to decide which. Numbering recalculated to O94868 magi1-fchsd2-5 1 7 unspecified role unspecified role prey prey pdz binding region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain in vitro In vitro 11 unspecified role unspecified role bait bait region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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) physical association physical association In the reverse experiment, we prepared 35S-methionine-labeled Carom by in vitro transcription translation, and performed pulldowns with GST proteins containing various PDZ domains of MAGI-1. Figure 3Bc From sequence length, FCHSD2 must be isoform -2 or -3 but not enough information given to decide which. Numbering recalculated to O94868 magi1-fchsd2-6 1 7 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 pdz region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined [EBI-1215916]Certain and ragged-n-terminus positions must always be strictly superior to 0 and inferior or equal to the protein sequence length. Actual positions : 975-975, sequence length 740 [EBI-1215916]975-1256 9 competitor competitor competitor prey prey camk region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 11 unspecified role unspecified role prey prey 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 association association How-ever, the addition of GST-CASK-Camk, but not GST inhibited the interaction between GST-MAGI-1-PDZ4+5 and GST-Carom-1 in a dose-dependent manner. 6 From sequence length, FCHSD2 must be isoform -2 or -3 but not enough information given to decide which. Numbering recalculated to O94868 fchsd2-cask-1 2 6 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association When the lysates of MDCK cells were incubated with the anti-Carom antibody, the immunoprecipitate contained CASK (Figure 5B). 5B cask-fchsd2-1 1 9 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 camk region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 7 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 physical association physical association We incubated various Myc-tagged proteins of CASK with either GST or GST-Carom-1 (Figure 5Ca). 5C From sequence length, FCHSD2 must be isoform -2 or -3 but not enough information given to decide which. Numbering recalculated to O94868 magi1-fchsd2-7 3 8 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 7 unspecified role unspecified role neutral component neutral component colocalization colocalization Then, cells started to contact with each other and GFP-Carom and MAGI-1 were accumulated at the cell contacts (Ca2+, 15 min). 7 Observed after 15 minutes re-exposure to calcium, not visible after 2hrs re-exposure. fchsd2-cask-2 3 6 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 7 unspecified role unspecified role neutral component neutral component physical association physical association CASK was also accumulated at cell contacts and colocalized with GFP-Carom (Figure 7Bb-2, arrows). 7