mbinfo MBInfo http://www.mechanobio.info/ http://www.mechanobio.info/ kurita-2011-1 Cooperative role of nectin-nectin and nectin-afadin interactions in formation of nectin-based cell-cell adhesion. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Kurita S., Ogita H., Takai Y. The source of GFP-afadin and Flag-Nectin 3 traced from references. ytakai@ med.kobe-u.ac.jp ytakai@med.kobe-u.ac.jp J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation kurita-2011-2 Cooperative role of nectin-nectin and nectin-afadin interactions in formation of nectin-based cell-cell adhesion. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Kurita S., Ogita H., Takai Y. The source of afadin (endogenous) inferred from the cells (HEK293) used. The source of Flag-Nectin 3 traced from references. The source of GFP-afadin and Flag-Nectin 3 traced from references. ytakai@ med.kobe-u.ac.jp ytakai@med.kobe-u.ac.jp J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation nect3_mouse Nectin-3 Nectin3 Nectin cell adhesion molecule 3 Poliovirus receptor-related protein 3 Pvrl3 protein protein mouse Mus musculus Mouse MARTPGPAPLCPGGGKAQLSSAFPPAAGLLLPAPTPPPLLLLLIPLLLFSRLCGALAGSIIVEPHVTAVWGKNVSLKCLIEVNETITQISWEKIHGKSTQTVAVHHPQYGFSVQGDYQGRVLFKNYSLNDATITLHNIGFSDSGKYICKAVTFPLGNAQSSTTVTVLVEPTVSLIKGPDSLIDGGNETVAAVCVAATGKPVAQIDWEGDLGEMESSTTSFPNETATIVSQYKLFPTRFARGRRITCVVKHPALEKDIRYSFILDIQYAPEVSVTGYDGNWFVGRKGVNLKCNADANPPPFKSVWSRLDGQWPDGLLASDNTLHFVHPLTVNYSGVYVCKVSNSLGQRSDQKVIYISDPPTTTTLQPTVQWHSSPADVQDIATEHKKLPFPLSTLATLKDDTIGTIIASVVGGALFLVLVSILAGVFCYRRRRTFRGDYFAKNYIPPSDMQKESQIDVLHQDELDSYPDSVKKENKNPVNNLIRKDYLEEPEKTQWNNVENLTRFERPMDYYEDLKMGMKFVSDERYNESEDGLVSHVDGSVISRREWYV afad_rat Afadin Afdn Af6 Afadin adherens junction formation factor Mllt4 Protein Af-6 protein protein rat Rattus norvegicus (Rat) Rat MSAGGRDEERRKLADIIHHWNANRLDLFEISQPTEDLEFHGVMRFYFQDKAAGNFATKCIRVSSTATTQDVIETLAEKFRPDMRMLSSPKYSLYEVHVSGERRLDIDEKPLVVQLNWNKDDREGRFVLKNENDAIPAKKAQSNGPEKQEKEGVIQNFKRTLSKKEKKEKKKREKEALRQASDKEERPSQGDDSENSRLAAEVYKDMPETSFTRTISNPEVVMKRRRQQKLEKRMQEFRSSDGRPDSGGTLRIYADSLKPNIPYKTILLSTTDPADFAVAESLEKYGLEKENPKDYCIARVMLPPGAQHSDERGAKEIILDDDECPLQIFREWPSDKGILVFQLKRRPPDYIPKKMKKHVEGKPLKGKDRADGSGYGSALPPEKLPYLVELSPGRRNHFAYYSYHTYEDGSDSRDKPKLYRLQLSVTEVGTEKFDDNSIQLFGPGIQPHHCDLTNMDGVVTVTPRSMDAETYVDGQRISETTMLQSGMRLQFGTSHVFKFVDPIQDHVLSKRSVDGGLMVKGPRHKPGAVQETTFELGGDIHSGTALPASRSTTRLDSDRVSSASSTAERGMVKPMIRLDQEQDYRRRESRTQDAAGPELMLPASIEFRESSEDSFLSAIINYTNSSTVHFKLSPTYVLYMACRYVLSSQHRPDISPTERTHKAIAVVNKMVSMMEGVIQEVDQVDQKQKNIAGALAFWMANASELLNFIKQDRDLSRITLDAQDVLAHLVQMAFKYLVHCLQSELNNYMPAFLDDPEENSLQRPKIDDVLHTLTGAMSLLRRCRVNAALTIQLFSQLFHFINMWLFNRLVTDPDSGLCSHYWGAIIRQQLGHIEAWAEKQGLELAADCHLSRIVQATTLLTMDKYVPDDIPNINSTCFKLNSLQLQALLQNYHCAPDEPFIPTDLIENVVAVAENTADELARSDGRDVQLEEDPDLQLPFLLPEDGYSCDVVRNIPNGLQEFLDPLCQRGFCRLVPHTRSPGTWTIYFEGADYESHLMRENTELTQPLRKEPEVITVTLKKQNGMGLSIVAAKGAGQDKLGIYVKSVVKGGAADVDGRLAAGDQLLSVDGRSLVGLSQERAAELMTRTSSVVTLEVAKQGAIYHGLATLLNQPSPMMQRISDRRGSGKPRPKSEGFELYNNSAQNGSPESPQMPWTEYSEPKKLPGDDRLMKNRADHRSSPNVANQPPSPGGKSPYTSGTAAKITSVSTGNLCTEEQTPPPRPEAYPIPTQTYTREYFTFPASKSQDRMAPVQNQWPNYEEKPHMHTESDHASIAIQRVTRSQEELREEKVYQLERHRVESGMDRKCDSDMWINQSSSVESSTSSQEHLNHSSKSVTPASTLTKSGPGRWKTPAAVLPTPVAVSQPIRTDLPPPPPPPPAHYTSDFDGISMDLPLPPPPANQAAPQSAQVAAAERKKREEHQRWYEKEKARLEEERERKRREQERKLGQMRTQSLNPASFSPLATQAKPEKPSTLQRPQETVIRELQPQQQPRTIERRDLQYITISKEELSSGDSLSPDPWKRDAREKLEKQQQMHIVDMLSKEIHELQNKGDRTAEESDRLRKLMLEWQFQKRLQESKQKDEDDDEEEDDDVDTMLIMQRLEAERRARLQDEERRRQQQLEEMRKREVEDRVRQEEDGRHQEEERVKRDAEEKRRQEEGYYSRLEAERRRQHEEAARRLLEPEEPGLSRPPLPQDYEPPSQSSAPSAPPPPPQRNASYLKTQVLSPDSLFTAKFVAYDDDDEEENYVPAGPNSYSGSAGTTAGTYDAPRDTREKLSRSQDADLPGSSGAPENLTFRERQRLFSQGQDVSDKVKASRKLTELENELNTK afad_human Afadin AFDN AF6 ALL1-fused gene from chromosome 6 protein Afadin adherens junction formation factor MLLT4 protein protein human Homo sapiens Human MSAGGRDEERRKLADIIHHWNANRLDLFEISQPTEDLEFHGVMRFYFQDKAAGNFATKCIRVSSTATTQDVIETLAEKFRPDMRMLSSPKYSLYEVHVSGERRLDIDEKPLVVQLNWNKDDREGRFVLKNENDAIPPKKAQSNGPEKQEKEGVIQNFKRTLSKKEKKEKKKREKEALRQASDKDDRPFQGEDVENSRLAAEVYKDMPETSFTRTISNPEVVMKRRRQQKLEKRMQEFRSSDGRPDSGGTLRIYADSLKPNIPYKTILLSTTDPADFAVAEALEKYGLEKENPKDYCIARVMLPPGAQHSDEKGAKEIILDDDECPLQIFREWPSDKGILVFQLKRRPPDHIPKKTKKHLEGKTPKGKERADGSGYGSTLPPEKLPYLVELSPGRRNHFAYYNYHTYEDGSDSRDKPKLYRLQLSVTEVGTEKLDDNSIQLFGPGIQPHHCDLTNMDGVVTVTPRSMDAETYVEGQRISETTMLQSGMKVQFGASHVFKFVDPSQDHALAKRSVDGGLMVKGPRHKPGIVQETTFDLGGDIHSGTALPTSKSTTRLDSDRVSSASSTAERGMVKPMIRVEQQPDYRRQESRTQDASGPELILPASIEFRESSEDSFLSAIINYTNSSTVHFKLSPTYVLYMACRYVLSNQYRPDISPTERTHKVIAVVNKMVSMMEGVIQKQKNIAGALAFWMANASELLNFIKQDRDLSRITLDAQDVLAHLVQMAFKYLVHCLQSELNNYMPAFLDDPEENSLQRPKIDDVLHTLTGAMSLLRRCRVNAALTIQLFSQLFHFINMWLFNRLVTDPDSGLCSHYWGAIIRQQLGHIEAWAEKQGLELAADCHLSRIVQATTLLTMDKYAPDDIPNINSTCFKLNSLQLQALLQNYHCAPDEPFIPTDLIENVVTVAENTADELARSDGREVQLEEDPDLQLPFLLPEDGYSCDVVRNIPNGLQEFLDPLCQRGFCRLIPHTRSPGTWTIYFEGADYESHLLRENTELAQPLRKEPEIITVTLKKQNGMGLSIVAAKGAGQDKLGIYVKSVVKGGAADVDGRLAAGDQLLSVDGRSLVGLSQERAAELMTRTSSVVTLEVAKQGAIYHGLATLLNQPSPMMQRISDRRGSGKPRPKSEGFELYNNSTQNGSPESPQLPWAEYSEPKKLPGDDRLMKNRADHRSSPNVANQPPSPGGKSAYASGTTAKITSVSTGNLCTEEQTPPPRPEAYPIPTQTYTREYFTFPASKSQDRMAPPQNQWPNYEEKPHMHTDSNHSSIAIQRVTRSQEELREDKAYQLERHRIEAAMDRKSDSDMWINQSSSLDSSTSSQEHLNHSSKSVTPASTLTKSGPGRWKTPAAIPATPVAVSQPIRTDLPPPPPPPPVHYAGDFDGMSMDLPLPPPPSANQIGLPSAQVAAAERRKREEHQRWYEKEKARLEEERERKRREQERKLGQMRTQSLNPAPFSPLTAQQMKPEKPSTLQRPQETVIRELQPQQQPRTIERRDLQYITVSKEELSSGDSLSPDPWKRDAKEKLEKQQQMHIVDMLSKEIQELQSKPDRSAEESDRLRKLMLEWQFQKRLQESKQKDEDDEEEEDDDVDTMLIMQRLEAERRARLQDEERRRQQQLEEMRKREAEDRARQEEERRRQEEERTKRDAEEKRRQEEGYYSRLEAERRRQHDEAARRLLEPEAPGLCRPPLPRDYEPPSPSPAPGAPPPPPQRNASYLKTQVLSPDSLFTAKFVAYNEEEEEEDCSLAGPNSYPGSTGAAVGAHDACRDAKEKRSKSQDADSPGSSGAPENLTFKERQRLFSQGQDVSNKVKASRKLTELENELNTK mllt4-pvrl3-6 1450152 n/a 1450154 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false Q9JLB9:p.Phe153Leu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false n/a 1450155 unspecified role unspecified role bait bait n_terminus gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false pdz domain required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig 1B, 2A, 2B, 3, 4, 5. The coprecipitation of Flag-Nectin 3 with GFP-afadin drastically reduced when the cells were not incubated with Fc-Nef-1 (fragment of mouse Nectin-1 fused to the Fc portion of human IgG). (Fig 1B, 2A, 2B, 3). Flag-Nectin 3 did not coprecipitate with GFP-afadin-deltaPDZ or deltaC constructs, in the presence or absence of Fc-Nef-1 (Fig 2). Deletion of the first Ig-like loop of nectin-3 reduced binding. Flag-Nectin 3 coprecipitated with GFP-afadin when the cells were incubated with Fc-Lef-5 (a fragment of mouse Necl-5 fused to the Fc portion of human IgG). Flag-Nectin 3 did not coprecipitate with GFP-afadin in the absence of Fc-Lef-5 (Fig 4). mllt4-pvrl3-5 1450153 n/a 1450156 unspecified role unspecified role bait bait n/a 1450154 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig 1C. The coprecipitation of Flag-Nectin 3 with afadin was significantly increased when the cells were incubated with Fc-Nef-1 (a fragment of mouse nectin-1 fused to the Fc portion of human IgG).