mbinfo MBInfo http://www.mechanobio.info/ Two cell adhesion molecules, nectin and cadherin, interact through their cytoplasmic domain-associated proteins. Two cell adhesion molecules, nectin and cadherin, interact through their cytoplasmic domain-associated proteins. J. Cell Biol. (0021-9525) 2000 imex curation Tachibana K., Nakanishi H., Mandai K., Ozaki K., Ikeda W., Yamamoto Y., Nagafuchi A., Tsukita S., Takai Y. The colocalization experiments (except the Nectin-2-l-Afadin colocalizations) were not curated, since there were no other experimental procedures demonstrating a physical interaction between these sets of proteins. ytakai@molbio.med.osaka-u.ac 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions mouse-f9 Mus musculus f9 Mouse F9 129 embryo teratocarcinoma confocal microscopy confocal microscopy immunostaining immunostaining Tachibana K., Nakanishi H., Mandai K., Ozaki K., Ikeda W., Yamamoto Y., Nagafuchi A., Tsukita S., Takai Y. The interaction detection and participant detection methods traced from references. The colocalization experiments (except the Nectin-2-l-Afadin colocalizations) were not curated, since there were no other experimental procedures demonstrating a physical interaction between these sets of proteins. ytakai@molbio.med.osaka-u.ac Accepted 2013-NOV-27 by ORCHARD J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation Two cell adhesion molecules, nectin and cadherin, interact through their cytoplasmic domain-associated proteins. Two cell adhesion molecules, nectin and cadherin, interact through their cytoplasmic domain-associated proteins. J. Cell Biol. (0021-9525) 2000 imex curation Tachibana K., Nakanishi H., Mandai K., Ozaki K., Ikeda W., Yamamoto Y., Nagafuchi A., Tsukita S., Takai Y. The colocalization experiments (except the Nectin-2-l-Afadin colocalizations) were not curated, since there were no other experimental procedures demonstrating a physical interaction between these sets of proteins. ytakai@molbio.med.osaka-u.ac 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions mouse-l cells Mus musculus embryo fibroblast, Moloney Sarcoma Virus transformed l cells Mouse embryo fibroblast, Moloney Sarcoma Virus transformed confocal microscopy confocal microscopy immunostaining immunostaining Tachibana K., Nakanishi H., Mandai K., Ozaki K., Ikeda W., Yamamoto Y., Nagafuchi A., Tsukita S., Takai Y. The interaction detection and participant detection methods traced from references. The colocalization experiments (except the Nectin-2-l-Afadin colocalizations) were not curated, since there were no other experimental procedures demonstrating a physical interaction between these sets of proteins. ytakai@molbio.med.osaka-u.ac Accepted 2013-NOV-27 by ORCHARD J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation Two cell adhesion molecules, nectin and cadherin, interact through their cytoplasmic domain-associated proteins. Two cell adhesion molecules, nectin and cadherin, interact through their cytoplasmic domain-associated proteins. J. Cell Biol. (0021-9525) 2000 imex curation Tachibana K., Nakanishi H., Mandai K., Ozaki K., Ikeda W., Yamamoto Y., Nagafuchi A., Tsukita S., Takai Y. The colocalization experiments (except the Nectin-2-l-Afadin colocalizations) were not curated, since there were no other experimental procedures demonstrating a physical interaction between these sets of proteins. ytakai@molbio.med.osaka-u.ac 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae 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 Tachibana K., Nakanishi H., Mandai K., Ozaki K., Ikeda W., Yamamoto Y., Nagafuchi A., Tsukita S., Takai Y. The colocalization experiments (except the Nectin-2-l-Afadin colocalizations) were not curated, since there were no other experimental procedures demonstrating a physical interaction between these sets of proteins. ytakai@molbio.med.osaka-u.ac Accepted 2013-NOV-27 by ORCHARD J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation Two cell adhesion molecules, nectin and cadherin, interact through their cytoplasmic domain-associated proteins. Two cell adhesion molecules, nectin and cadherin, interact through their cytoplasmic domain-associated proteins. J. Cell Biol. (0021-9525) 2000 imex curation Tachibana K., Nakanishi H., Mandai K., Ozaki K., Ikeda W., Yamamoto Y., Nagafuchi A., Tsukita S., Takai Y. The colocalization experiments (except the Nectin-2-l-Afadin colocalizations) were not curated, since there were no other experimental procedures demonstrating a physical interaction between these sets of proteins. ytakai@molbio.med.osaka-u.ac 13-Jan-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Tachibana K., Nakanishi H., Mandai K., Ozaki K., Ikeda W., Yamamoto Y., Nagafuchi A., Tsukita S., Takai Y. interaction detection method: Myc-His6 pulldown using metal affinity beads. The colocalization experiments (except the Nectin-2-l-Afadin colocalizations) were not curated, since there were no other experimental procedures demonstrating a physical interaction between these sets of proteins. ytakai@molbio.med.osaka-u.ac Accepted 2013-NOV-27 by ORCHARD J. Cell Biol. (0021-9525) 2000 Only protein-protein interactions imex curation ctna1_mouse Catenin alpha-1 102 kDa cadherin-associated protein CAP102 Alpha E-catenin Ctnna1 Catna1 protein protein mouse Mus musculus Mouse MTAVHAGNINFKWDPKSLEIRTLAVERLLEPLVTQVTTLVNTNSKGPSNKKRGRSKKAHVLAASVEQATENFLEKGDKIAKESQFLKEELVVAVEDVRKQGDLMKSAAGEFADDPCSSVKRGNMVRAARALLSAVTRLLILADMADVYKLLVQLKVVEDGILKLRNAGNEQDLGIQYKALKPEVDKLNIMAAKRQQELKDVGNRDQMAAARGILQKNVPILYTASQACLQHPDVAAYKANRDLIYKQLQQAVTGISNAAQATASDDAAQHQGGSGGELAYALNNFDKQIIVDPLSFSEERFRPSLEERLESIISGAALMADSSCTRDDRRERIVAECNAVRQALQDLLSEYMGNAGRKERSDALNSAIDKMTKKTRDLRRQLRKAVMDHVSDSFLETNVPLLVLIEAAKNGNEKEVKEYAQVFREHANKLIEVANLACSISNNEEGVKLVRMSASQLEALCPQVINAALALAAKPQSKLAQENMDLFKEQWEKQVRVLTDAVDDITSIDDFLAVSENHILEDVNKCVIALQEKDVDGLDRTAGAIRGRAARVIHVVTSEMDNYEPGVYTEKVLEATKLLSNTVMPRFTEQVEAAVEALSSDPAQPMDENEFIDASRLVYDGIRDIRKAVLMIRTPEELDDSDFETEDFDVRSRTSVQTEDDQLIAGQSARAIMAQLPQEQKAKIAEQVASFQEEKSKLDAEVSKWDDSGNDIIVLAKQMCMIMMEMTDFTRGKGPLKNTSDVISAAKKIAEAGSRMDKLGRTIADHCPDSACKQDLLAYLQRIALYCHQLNICSKVKAEVQNLGGELVVSGVDSAMSLIQAAKNLMNAVVQTVKASYVASTKYQKSQGMASLNLPAVSWKMKAPEKKPLVKREKQDETQTKIKRASQKKHVNPVQALSEFKAMDSI DIP protein P26231 original sequence version: 125 DIP protein P26231 original sequence version: 142 DIP protein P26231 original sequence version: 160 3ED1EC50925DBBB5 q9qzq1-2 Afadin Afdn Af6 Afadin adherens junction formation factor Mllt4 Protein Af-6 l-afadin protein protein mouse Mus musculus Mouse MSAGGRDEERRKLADIIHHWNANRLDLFEISQPTEDLEFHGVMRFYFQDKAAGNFATKCIRVSSTATTQDVIETLAEKFRPDMRMLSSPKYSLYEVHVSGERRLDIDEKPLVVQLNWNKDDREGRFVLKNENDAIPAKKAQSNGPEKQEKEGVIQNFKRTLSKKEKKEKKKKEKEALRQASDKEERPSQGDDSENSRLAAEVYKDMPETSFTRTISNPEVVMKRRRQQKLEKRMQEFRSSDGRPDSGGTLRIYADSLKPNIPYKTILLSTTDTADFAVAESLEKYGLEKENPKDYCIARVMLPPGAQHSDERGAKEIILDDDECPLQIFREWPSDKGILVFQLKRRPPDYIPKKMKKHVEGKSLKGKDRADGSGYGSALPPEKLPYLVELSPDGSDSRDKPKLYRLQLSVTEVGTEKFDDNSIQLFGPGIQPHHCDLTNMDGVVTVTPRSMDAETYVDGQRISETTMLQSGMRLQFGTSHVFKFVDPIQDHVLSKRSVDGGLMVKGPRHKPGAVQETTFELGGDVHSGTALPASRSTTRLDSDRVSSASSTAERGMVKPMIRLDQEQEYRRRENRTQDATGPELILPASIEFRESSEDSFLSAIINYTNSSTVHFKLSPTYVLYMACRYVLSSQHRPDISPTERTHKAIAVVNKMVSMMEGVIQKQKNIAGALAFWMANASELLNFIKQDRDLSRITLDAQDVLAHLVQMAFKYLVHCLQSELNNYMPAFLDDPEENSLQRPKIDDVLHTLTGAMSLLRRCRVNAALTIQLFSQLFHFINMWLFNRLVTDPDSGLCSHYWGAIIRQQLGHIEAWAEKQGLELAADCHLSRIVQATTLLTMDKYVPDDIPNINSTCFKLNSLQLQALLQNYHCAPDEPFIPTDLIENVVAVAENTADELARSDGRDVQLEEDPDLQLPFLLPEDGYSCDVVRNIPNGLQEFLDPLCQRGFCRLVPHTRSPGTWTIYFEGADYESHLMRENAELAQPLRKEPEIITVTLKKQNGMGLSIVAAKGAGQDKLGIYVKSVVKGGAADVDGRLAAGDQLLSVDGRSLVGLSQERAAELMTRTSSVVTLEVAKQGAIYHGLATLLNQPSPMMQRISDRRGSGKPRPKSEGFELYNNSAQNGSPESPQMPWTEYSEPKKLPGDDRLMKNRADHRSSPNVANQPPSPGGKGPYTSGTAAKITSVSTGNLCTEEQSPPPRPEAYPIPTQTYTREYFTFPASKSQDRMAPPQSQWPNYEEKPHVHTESNHSSIAIQRVTRSQEELREEKVYQLERHRVEAGMDRKCDSDMWINQSSSVESSTSSQEHLNHSSKSVTPASTLTKSGPGRWKTPAAVLPTPVAVSQPIRTDLPPPPPPPPVHYTSEFDGIPMDLPLPPPPANQAGPQSAQVAAAEWKKREEHQRWYEKEKARLEEERERKRREQERKLGQMRSQTLNPASFSPLATQAKPEKPSTLQRPQETVIRELQPQQQPRTIERKDLQYITISKEELSSGDSLSPDPWKRDAREKLEKQQQMHIVDMLSKEIHELQNKVDRTAEESDRLRKLMLEWQFQKRLQESKQKDEDDDEEEDDDVDTMLIMQRLEAERRARMQDEERRRQQQLEEMRKREAEDRVRQEEDGRHQEEERVKRDAEEKRRQEEGYYSRLEAERRRQHEEAARRLLEPEEPGLSRPPLPRDYEPPSLSSAPCAPPPPPQRNASYLKTQVLSPDSLFTAKFVAYDEEEEDYGPAGPNSYSGSAGTAVGAYDAPREAREKLTRSQDADLPGSSGAPENLTFKERQRLFSQGQDVSDKVKASRKLTELENELNTK 5987A16BA9F28C2E afad_rat Afadin Afdn Af6 Afadin adherens junction formation factor Mllt4 Protein Af-6 protein protein rat Rattus norvegicus (Rat) Rat MSAGGRDEERRKLADIIHHWNANRLDLFEISQPTEDLEFHGVMRFYFQDKAAGNFATKCIRVSSTATTQDVIETLAEKFRPDMRMLSSPKYSLYEVHVSGERRLDIDEKPLVVQLNWNKDDREGRFVLKNENDAIPAKKAQSNGPEKQEKEGVIQNFKRTLSKKEKKEKKKREKEALRQASDKEERPSQGDDSENSRLAAEVYKDMPETSFTRTISNPEVVMKRRRQQKLEKRMQEFRSSDGRPDSGGTLRIYADSLKPNIPYKTILLSTTDPADFAVAESLEKYGLEKENPKDYCIARVMLPPGAQHSDERGAKEIILDDDECPLQIFREWPSDKGILVFQLKRRPPDYIPKKMKKHVEGKPLKGKDRADGSGYGSALPPEKLPYLVELSPGRRNHFAYYSYHTYEDGSDSRDKPKLYRLQLSVTEVGTEKFDDNSIQLFGPGIQPHHCDLTNMDGVVTVTPRSMDAETYVDGQRISETTMLQSGMRLQFGTSHVFKFVDPIQDHVLSKRSVDGGLMVKGPRHKPGAVQETTFELGGDIHSGTALPASRSTTRLDSDRVSSASSTAERGMVKPMIRLDQEQDYRRRESRTQDAAGPELMLPASIEFRESSEDSFLSAIINYTNSSTVHFKLSPTYVLYMACRYVLSSQHRPDISPTERTHKAIAVVNKMVSMMEGVIQEVDQVDQKQKNIAGALAFWMANASELLNFIKQDRDLSRITLDAQDVLAHLVQMAFKYLVHCLQSELNNYMPAFLDDPEENSLQRPKIDDVLHTLTGAMSLLRRCRVNAALTIQLFSQLFHFINMWLFNRLVTDPDSGLCSHYWGAIIRQQLGHIEAWAEKQGLELAADCHLSRIVQATTLLTMDKYVPDDIPNINSTCFKLNSLQLQALLQNYHCAPDEPFIPTDLIENVVAVAENTADELARSDGRDVQLEEDPDLQLPFLLPEDGYSCDVVRNIPNGLQEFLDPLCQRGFCRLVPHTRSPGTWTIYFEGADYESHLMRENTELTQPLRKEPEVITVTLKKQNGMGLSIVAAKGAGQDKLGIYVKSVVKGGAADVDGRLAAGDQLLSVDGRSLVGLSQERAAELMTRTSSVVTLEVAKQGAIYHGLATLLNQPSPMMQRISDRRGSGKPRPKSEGFELYNNSAQNGSPESPQMPWTEYSEPKKLPGDDRLMKNRADHRSSPNVANQPPSPGGKSPYTSGTAAKITSVSTGNLCTEEQTPPPRPEAYPIPTQTYTREYFTFPASKSQDRMAPVQNQWPNYEEKPHMHTESDHASIAIQRVTRSQEELREEKVYQLERHRVESGMDRKCDSDMWINQSSSVESSTSSQEHLNHSSKSVTPASTLTKSGPGRWKTPAAVLPTPVAVSQPIRTDLPPPPPPPPAHYTSDFDGISMDLPLPPPPANQAAPQSAQVAAAERKKREEHQRWYEKEKARLEEERERKRREQERKLGQMRTQSLNPASFSPLATQAKPEKPSTLQRPQETVIRELQPQQQPRTIERRDLQYITISKEELSSGDSLSPDPWKRDAREKLEKQQQMHIVDMLSKEIHELQNKGDRTAEESDRLRKLMLEWQFQKRLQESKQKDEDDDEEEDDDVDTMLIMQRLEAERRARLQDEERRRQQQLEEMRKREVEDRVRQEEDGRHQEEERVKRDAEEKRRQEEGYYSRLEAERRRQHEEAARRLLEPEEPGLSRPPLPQDYEPPSQSSAPSAPPPPPQRNASYLKTQVLSPDSLFTAKFVAYDDDDEEENYVPAGPNSYSGSAGTTAGTYDAPRDTREKLSRSQDADLPGSSGAPENLTFRERQRLFSQGQDVSDKVKASRKLTELENELNTK 45C597A82F109D6F p32507-2 Nectin-2 Nectin2 Herpes virus entry mediator B Mph Murine herpes virus entry protein B Nectin cell adhesion molecule 2 Poliovirus receptor homolog Poliovirus receptor-related protein 2 Pvr Pvrl2 Pvs protein protein mouse Mus musculus Mouse MARAAVLPPSRLSPTLPLLPLLLLLLQETGAQDVRVRVLPEVRGRLGGTVELPCHLLPPTTERVSQVTWQRLDGTVVAAFHPSFGVDFPNSQFSKDRLSFVRARPETNADLRDATLAFRGLRVEDEGNYTCEFATFPNGTRRGVTWLRVIAQPENHAEAQEVTIGPQSVAVARCVSTGGRPPARITWISSLGGEAKDTQEPGIQAGTVTIISRYSLVPVGRADGVKVTCRVEHESFEEPILLPVTLSVRYPPEVSISGYDDNWYLGRSEAILTCDVRSNPEPTDYDWSTTSGVFPASAVAQGSQLLVHSVDRMVNTTFICTATNAVGTGRAEQVILVRDTPQASRDVGPLVWGAVGGTLLVLLLAGGFLALILLRGRRRRKSPGGGGNDGDRGSYDPKTQVFGNGGPVFWRSASPEPMRPDGREEDEEEEEEMKAEEGLMLPPHESPKDDMESHLDGSLISRRAVYV DIP protein P32507-2 original sequence version: 114 7DB2430F6FFDD333 nect2_mouse Nectin-2 Nectin2 Herpes virus entry mediator B Mph Murine herpes virus entry protein B Nectin cell adhesion molecule 2 Poliovirus receptor homolog Poliovirus receptor-related protein 2 Pvr Pvrl2 Pvs protein protein mouse Mus musculus Mouse MARAAVLPPSRLSPTLPLLPLLLLLLQETGAQDVRVRVLPEVRGRLGGTVELPCHLLPPTTERVSQVTWQRLDGTVVAAFHPSFGVDFPNSQFSKDRLSFVRARPETNADLRDATLAFRGLRVEDEGNYTCEFATFPNGTRRGVTWLRVIAQPENHAEAQEVTIGPQSVAVARCVSTGGRPPARITWISSLGGEAKDTQEPGIQAGTVTIISRYSLVPVGRADGVKVTCRVEHESFEEPILLPVTLSVRYPPEVSISGYDDNWYLGRSEAILTCDVRSNPEPTDYDWSTTSGVFPASAVAQGSQLLVHSVDRMVNTTFICTATNAVGTGRAEQVILVRESPSTAGAGATGGIIGGIIAAIIATAVAGTGILICRQQRKEQRLQAADEEEELEGPPSYKPPTPKAKLEEPEMPSQLFTLGASEHSPVKTPYFDAGVSCADQEMPRYHELPTLEERSGPLLLGATGLGPSLLVPPGPNVVEGVSLSLEDEEEDDEEEDFLDKINPIYDALSYPSPSDSYQSKDFFVSRAMYV 0ED71BFA2B231BBE ctnna1-mllt4-1 3 5 unspecified role unspecified role bait bait c-terminal region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain n-terminal region 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 7 unspecified role unspecified role prey prey n-terminal region 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 physical association physical association Table 1. pvrl2-mllt4-5 3 8 unspecified role unspecified role bait bait n-terminal region 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 cytoplasmic region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role prey prey n-terminal region 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 physical association physical association Table 1. mllt4-ctnna1-1 4 5 unspecified role unspecified role prey prey n-terminal region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range c-terminal 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) 7 unspecified role unspecified role bait bait c-terminal region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range c-terminal region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. physical association physical association Fig 6. mllt4-pvrl2-1 1 9 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig 2A. Nectin-2 and l-Afadin colocalized at cell-cell adhesion sites only in cells transfected with alpha-catenin and not in untransfected cells (Fig 2A, 2B). mllt4-pvrl2-3 2 9 unspecified role unspecified role neutral component neutral component 6 unspecified role unspecified role neutral component neutral component colocalization colocalization Fig 3A, 5A, 7. Nectin-2 colocalized with l-Afadin at cell-cell adhesion sites even in the absence of beta-catenin. ie., in cells transfected with a chimeric protein, where an E-cadherin lacking the beta-catenin binding domain is fused to full length alpha-catenin (Fig 3A). Nectin-2 colocalized with l-Afadin at cell-cell adhesion sites even in the absence of vinculin and ponsin, ie., in cells transfected with a chimeric protein, where E-cadherin is fused to the C-terminal region of alpha-catenin, which lacks binding to vinculin and ponsin (Fig 5A). l-Afadin colocalized with Nectin-2 at cell-cell adhesion sites in cells transfected with full length Nectin-2alpha, but not in cells transfected with mutant Nectin-2alpha with the last the last 4 carboxyl terminus residues deleted (Fig 7).