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/ http://www.ebi.ac.uk/ rantala-2011-7 SHARPIN is an endogenous inhibitor of β1-integrin activation. human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Rantala J.K., Pouwels J., Pellinen T., Veltel S., Laasola P., Mattila E., Potter C.S., Duffy T., Sundberg J.P., Kallioniemi O., Askari J.A., Humphries M.J., Parsons M., Salmi M., Ivaska J. Species of alpha5-gfp, mcherry-kindlin amd mcherry-talin confirmed by the authors. Figure 4d (PLA) has not been included as the identity of the target Kindlin molecule(s) is not known. Johanna.ivaska@vtt.fi Nat. Cell Biol. (1465-7392) 2011 Only protein-protein interactions imex curation rantala-2011-1 SHARPIN is an endogenous inhibitor of β1-integrin activation. human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma pull down pull down anti tag western anti tag western blot anti tag western Rantala J.K., Pouwels J., Pellinen T., Veltel S., Laasola P., Mattila E., Potter C.S., Duffy T., Sundberg J.P., Kallioniemi O., Askari J.A., Humphries M.J., Parsons M., Salmi M., Ivaska J. Species of alpha5-gfp, mcherry-kindlin amd mcherry-talin confirmed by the authors. Figure 4d (PLA) has not been included as the identity of the target Kindlin molecule(s) is not known. Johanna.ivaska@vtt.fi Nat. Cell Biol. (1465-7392) 2011 Only protein-protein interactions imex curation rantala-2011-3 SHARPIN is an endogenous inhibitor of β1-integrin activation. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Rantala J.K., Pouwels J., Pellinen T., Veltel S., Laasola P., Mattila E., Potter C.S., Duffy T., Sundberg J.P., Kallioniemi O., Askari J.A., Humphries M.J., Parsons M., Salmi M., Ivaska J. Species of alpha5-gfp, mcherry-kindlin amd mcherry-talin confirmed by the authors. Figure 4d (PLA) has not been included as the identity of the target Kindlin molecule(s) is not known. Johanna.ivaska@vtt.fi Nat. Cell Biol. (1465-7392) 2011 Only protein-protein interactions imex curation rantala-2011-5 SHARPIN is an endogenous inhibitor of β1-integrin activation. mouse-gd25 Mouse GD25 embryonic fibroblasts, integrin beta1 null. gd25 Mouse GD25 embryonic fibroblasts, integrin beta1 null. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Rantala J.K., Pouwels J., Pellinen T., Veltel S., Laasola P., Mattila E., Potter C.S., Duffy T., Sundberg J.P., Kallioniemi O., Askari J.A., Humphries M.J., Parsons M., Salmi M., Ivaska J. Species of alpha5-gfp, mcherry-kindlin amd mcherry-talin confirmed by the authors. Figure 4d (PLA) has not been included as the identity of the target Kindlin molecule(s) is not known. Johanna.ivaska@vtt.fi Nat. Cell Biol. (1465-7392) 2011 Only protein-protein interactions imex curation rantala-2011-11 SHARPIN is an endogenous inhibitor of β1-integrin activation. mouse-embryonic_fibroblast Mus musculus embryonic fibroblasts embryonic_fibroblast Embryonic fibroblast Fibroblasts taken from an embryonic as opposed to a foetal or adult source. Foetus is defined as the stage at which tissue diferentiation is initiated. Tissue taken from an embryonic as opposed to a foetal or adult source. Embryo is defined as the development stage prior to organ formation. fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Rantala J.K., Pouwels J., Pellinen T., Veltel S., Laasola P., Mattila E., Potter C.S., Duffy T., Sundberg J.P., Kallioniemi O., Askari J.A., Humphries M.J., Parsons M., Salmi M., Ivaska J. Integrin-beta1 knockout mefs. Species of alpha5-gfp, mcherry-kindlin amd mcherry-talin confirmed by the authors. Figure 4d (PLA) has not been included as the identity of the target Kindlin molecule(s) is not known. Johanna.ivaska@vtt.fi Nat. Cell Biol. (1465-7392) 2011 Only protein-protein interactions imex curation rantala-2011-2 SHARPIN is an endogenous inhibitor of β1-integrin activation. human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma pull down pull down western blot western blot Immuno blot Immuno blot Rantala J.K., Pouwels J., Pellinen T., Veltel S., Laasola P., Mattila E., Potter C.S., Duffy T., Sundberg J.P., Kallioniemi O., Askari J.A., Humphries M.J., Parsons M., Salmi M., Ivaska J. Species of alpha5-gfp, mcherry-kindlin amd mcherry-talin confirmed by the authors. Figure 4d (PLA) has not been included as the identity of the target Kindlin molecule(s) is not known. Johanna.ivaska@vtt.fi Nat. Cell Biol. (1465-7392) 2011 Only protein-protein interactions imex curation rantala-2011-4 SHARPIN is an endogenous inhibitor of β1-integrin activation. in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps predetermined predetermined participant predetermined Rantala J.K., Pouwels J., Pellinen T., Veltel S., Laasola P., Mattila E., Potter C.S., Duffy T., Sundberg J.P., Kallioniemi O., Askari J.A., Humphries M.J., Parsons M., Salmi M., Ivaska J. Species of alpha5-gfp, mcherry-kindlin amd mcherry-talin confirmed by the authors. Figure 4d (PLA) has not been included as the identity of the target Kindlin molecule(s) is not known. Johanna.ivaska@vtt.fi Nat. Cell Biol. (1465-7392) 2011 Only protein-protein interactions imex curation rantala-2011-6 SHARPIN is an endogenous inhibitor of β1-integrin activation. human-nci_h460 Human NCI-H460 large cell lung carcinoma cell line nci_h460 Human NCI-H460 large cell lung carcinoma cell line ATCC =HTB-177 http://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=HTB-177&Template=cellBiology confocal microscopy confocal microscopy immunostaining immunostaining Rantala J.K., Pouwels J., Pellinen T., Veltel S., Laasola P., Mattila E., Potter C.S., Duffy T., Sundberg J.P., Kallioniemi O., Askari J.A., Humphries M.J., Parsons M., Salmi M., Ivaska J. Species of alpha5-gfp, mcherry-kindlin amd mcherry-talin confirmed by the authors. Figure 4d (PLA) has not been included as the identity of the target Kindlin molecule(s) is not known. Johanna.ivaska@vtt.fi Nat. Cell Biol. (1465-7392) 2011 Only protein-protein interactions imex curation rantala-2011-8 SHARPIN is an endogenous inhibitor of β1-integrin activation. human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Rantala J.K., Pouwels J., Pellinen T., Veltel S., Laasola P., Mattila E., Potter C.S., Duffy T., Sundberg J.P., Kallioniemi O., Askari J.A., Humphries M.J., Parsons M., Salmi M., Ivaska J. Species of alpha5-gfp, mcherry-kindlin amd mcherry-talin confirmed by the authors. Figure 4d (PLA) has not been included as the identity of the target Kindlin molecule(s) is not known. Johanna.ivaska@vtt.fi Nat. Cell Biol. (1465-7392) 2011 Only protein-protein interactions imex curation rantala-2011-9 SHARPIN is an endogenous inhibitor of β1-integrin activation. human-pc3 Human PC3 Caucasian prostate adenocarcinoma pc3 Human Caucasian prostate adenocarcinoma fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret tag fluorescence tag visualisation by fluorescence tag fluorescence Rantala J.K., Pouwels J., Pellinen T., Veltel S., Laasola P., Mattila E., Potter C.S., Duffy T., Sundberg J.P., Kallioniemi O., Askari J.A., Humphries M.J., Parsons M., Salmi M., Ivaska J. Species of alpha5-gfp, mcherry-kindlin amd mcherry-talin confirmed by the authors. Figure 4d (PLA) has not been included as the identity of the target Kindlin molecule(s) is not known. Johanna.ivaska@vtt.fi Nat. Cell Biol. (1465-7392) 2011 Only protein-protein interactions imex curation itb1_human Integrin beta-1 Fibronectin receptor subunit beta VLA-4 subunit beta ITGB1 FNRB MDF2 MSK12 Glycoprotein IIa protein protein human Homo sapiens Human MNLQPIFWIGLISSVCCVFAQTDENRCLKANAKSCGECIQAGPNCGWCTNSTFLQEGMPTSARCDDLEALKKKGCPPDDIENPRGSKDIKKNKNVTNRSKGTAEKLKPEDITQIQPQQLVLRLRSGEPQTFTLKFKRAEDYPIDLYYLMDLSYSMKDDLENVKSLGTDLMNEMRRITSDFRIGFGSFVEKTVMPYISTTPAKLRNPCTSEQNCTSPFSYKNVLSLTNKGEVFNELVGKQRISGNLDSPEGGFDAIMQVAVCGSLIGWRNVTRLLVFSTDAGFHFAGDGKLGGIVLPNDGQCHLENNMYTMSHYYDYPSIAHLVQKLSENNIQTIFAVTEEFQPVYKELKNLIPKSAVGTLSANSSNVIQLIIDAYNSLSSEVILENGKLSEGVTISYKSYCKNGVNGTGENGRKCSNISIGDEVQFEISITSNKCPKKDSDSFKIRPLGFTEEVEVILQYICECECQSEGIPESPKCHEGNGTFECGACRCNEGRVGRHCECSTDEVNSEDMDAYCRKENSSEICSNNGECVCGQCVCRKRDNTNEIYSGKFCECDNFNCDRSNGLICGGNGVCKCRVCECNPNYTGSACDCSLDTSTCEASNGQICNGRGICECGVCKCTDPKFQGQTCEMCQTCLGVCAEHKECVQCRAFNKGEKKDTCTQECSYFNITKVESRDKLPQPVQPDPVSHCKEKDVDDCWFYFTYSVNGNNEVMVHVVENPECPTGPDIIPIVAGVVAGIVLIGLALLLIWKLLMIIHDRREFAKFEKEKMNAKWDTGENPIYKSAVTTVVNPKYEGK ita2_human Integrin alpha-2 Platelet membrane glycoprotein Ia Collagen receptor VLA-2 subunit alpha CD49 antigen-like family member B ITGA2 CD49B protein protein human Homo sapiens Human MGPERTGAAPLPLLLVLALSQGILNCCLAYNVGLPEAKIFSGPSSEQFGYAVQQFINPKGNWLLVGSPWSGFPENRMGDVYKCPVDLSTATCEKLNLQTSTSIPNVTEMKTNMSLGLILTRNMGTGGFLTCGPLWAQQCGNQYYTTGVCSDISPDFQLSASFSPATQPCPSLIDVVVVCDESNSIYPWDAVKNFLEKFVQGLDIGPTKTQVGLIQYANNPRVVFNLNTYKTKEEMIVATSQTSQYGGDLTNTFGAIQYARKYAYSAASGGRRSATKVMVVVTDGESHDGSMLKAVIDQCNHDNILRFGIAVLGYLNRNALDTKNLIKEIKAIASIPTERYFFNVSDEAALLEKAGTLGEQIFSIEGTVQGGDNFQMEMSQVGFSADYSSQNDILMLGAVGAFGWSGTIVQKTSHGHLIFPKQAFDQILQDRNHSSYLGYSVAAISTGESTHFVAGAPRANYTGQIVLYSVNENGNITVIQAHRGDQIGSYFGSVLCSVDVDKDTITDVLLVGAPMYMSDLKKEEGRVYLFTIKKGILGQHQFLEGPEGIENTRFGSAIAALSDINMDGFNDVIVGSPLENQNSGAVYIYNGHQGTIRTKYSQKILGSDGAFRSHLQYFGRSLDGYGDLNGDSITDVSIGAFGQVVQLWSQSIADVAIEASFTPEKITLVNKNAQIILKLCFSAKFRPTKQNNQVAIVYNITLDADGFSSRVTSRGLFKENNERCLQKNMVVNQAQSCPEHIIYIQEPSDVVNSLDLRVDISLENPGTSPALEAYSETAKVFSIPFHKDCGEDGLCISDLVLDVRQIPAAQEQPFIVSNQNKRLTFSVTLKNKRESAYNTGIVVDFSENLFFASFSLPVDGTEVTCQVAASQKSVACDVGYPALKREQQVTFTINFDFNLQNLQNQASLSFQALSESQEENKADNLVNLKIPLLYDAEIHLTRSTNINFYEISSDGNVPSIVHSFEDVGPKFIFSLKVTTGSVPVSMATVIIHIPQYTKEKNPLMYLTGVQTDKAGDISCNADINPLKIGQTSSSVSFKSENFRHTKELNCRTASCSNVTCWLKDVHMKGEYFVNVTTRIWNGTFASSTFQTVQLTAAAEINTYNPEIYVIEDNTVTIPLMIMKPDEKAEVPTGVIIGSIIAGILLLLALVAILWKLGFFKRKYEKMTKNPDEIDETTELSS shrpn_human Sharpin SHARPIN SIPL1 Shank-associated RH domain-interacting protein Shank-interacting protein-like 1 PSEC0216 protein protein human Homo sapiens Human MAPPAGGAAAAASDLGSAAVLLAVHAAVRPLGAGPDAEAQLRRLQLSADPERPGRFRLELLGAGPGAVNLEWPLESVSYTIRGPTQHELQPPPGGPGTLSLHFLNPQEAQRWAVLVRGATVEGQNGSKSNSPPALGPEACPVSLPSPPEASTLKGPPPEADLPRSPGNLTEREELAGSLARAIAGGDEKGAAQVAAVLAQHRVALSVQLQEACFPPGPIRLQVTLEDAASAASAASSAHVALQVHPHCTVAALQEQVFSELGFPPAVQRWVIGRCLCVPERSLASYGVRQDGDPAFLYLLSAPREAPATGPSPQHPQKMDGELGRLFPPSLGLPPGPQPAASSLPSPLQPSWSCPSCTFINAPDRPGCEMCSTQRPCTWDPLAAAST ita5_human Integrin alpha-5 ITGA5 FNRA Fibronectin receptor subunit alpha Integrin alpha-F VLA-5 CD49 antigen-like family member E protein protein human Homo sapiens Human MGSRTPESPLHAVQLRWGPRRRPPLLPLLLLLLPPPPRVGGFNLDAEAPAVLSGPPGSFFGFSVEFYRPGTDGVSVLVGAPKANTSQPGVLQGGAVYLCPWGASPTQCTPIEFDSKGSRLLESSLSSSEGEEPVEYKSLQWFGATVRAHGSSILACAPLYSWRTEKEPLSDPVGTCYLSTDNFTRILEYAPCRSDFSWAAGQGYCQGGFSAEFTKTGRVVLGGPGSYFWQGQILSATQEQIAESYYPEYLINLVQGQLQTRQASSIYDDSYLGYSVAVGEFSGDDTEDFVAGVPKGNLTYGYVTILNGSDIRSLYNFSGEQMASYFGYAVAATDVNGDGLDDLLVGAPLLMDRTPDGRPQEVGRVYVYLQHPAGIEPTPTLTLTGHDEFGRFGSSLTPLGDLDQDGYNDVAIGAPFGGETQQGVVFVFPGGPGGLGSKPSQVLQPLWAASHTPDFFGSALRGGRDLDGNGYPDLIVGSFGVDKAVVYRGRPIVSASASLTIFPAMFNPEERSCSLEGNPVACINLSFCLNASGKHVADSIGFTVELQLDWQKQKGGVRRALFLASRQATLTQTLLIQNGAREDCREMKIYLRNESEFRDKLSPIHIALNFSLDPQAPVDSHGLRPALHYQSKSRIEDKAQILLDCGEDNICVPDLQLEVFGEQNHVYLGDKNALNLTFHAQNVGEGGAYEAELRVTAPPEAEYSGLVRHPGNFSSLSCDYFAVNQSRLLVCDLGNPMKAGASLWGGLRFTVPHLRDTKKTIQFDFQILSKNLNNSQSDVVSFRLSVEAQAQVTLNGVSKPEAVLFPVSDWHPRDQPQKEEDLGPAVHHVYELINQGPSSISQGVLELSCPQALEGQQLLYVTRVTGLNCTTNHPINPKGLELDPEGSLHHQQKREAPSRSSASSGPQILKCPEAECFRLRCELGPLHQQESQSLQLHFRVWAKTFLQREHQPFSLQCEAVYKALKMPYRILPRQLPQKERQVATAVQWTKAEGSYGVPLWIIILAILFGLLLLGLLIYILYKLGFFKRSLPYGTAMEKAQLKPPATSDA il1a_human Interleukin-1 alpha IL1A IL1F1 Hematopoietin-1 protein protein human Homo sapiens Human MAKVPDMFEDLKNCYSENEEDSSSIDHLSLNQKSFYHVSYGPLHEGCMDQSVSLSISETSKTSKLTFKESMVVVATNGKVLKKRRLSLSQSITDDDLEAIANDSEEEIIKPRSAPFSFLSNVKYNFMRIIKYEFILNDALNQSIIRANDQYLTAAALHNLDEAVKFDMGAYKSSKDDAKITVILRISKTQLYVTAQDEDQPVLLKEMPEIPKTITGSETNLLFFWETHGTKNYFTSVAHPNLFIATKQDYWVCLAGGPPSITDFQILENQA shrpn_rat Sharpin Sharpin Shank-associated RH domain-interacting protein protein protein rat Rattus norvegicus (Rat) Rat MSPPAGGAAAAADPASPVVLLAVQAAVRLLGAGHEDEAQLRKLQLKADPERPGRFRLGLLGIEPGAVSLEWPLESICYTIRGPNQHELQPPPGGPGTFSVHFLNSEEAQQWAALVRDATAEGQNGNDSTAPVPTPAMCPTSPPCSSVTPTPKATQPEMDLPQGSGNLKKEELATHLAQAIAGGDEKAAAQVAAILAQHHVALNVQLLEAWFPRGPIRLQVTVEDATSVLSSSSSAHVSLQIHPHCSIAALQEQVFSEFGFPPAVQRWVIGRCLCMPERSLASYGVSQDGDPAFLYLLSAPREVSGHSPQHSKMDRKLGCLFPQSLELPHNLQASSSSLPSPPQPGWSCPSCTFINASNRPGCEMCSTQRPCAWDPLTATST ita1_human Integrin alpha-1 ITGA1 Laminin and collagen receptor VLA-1 CD49 antigen-like family member A protein protein human Homo sapiens Human MAPRPRARPGVAVACCWLLTVVLRCCVSFNVDVKNSMTFSGPVEDMFGYTVQQYENEEGKWVLIGSPLVGQPKNRTGDVYKCPVGRGESLPCVKLDLPVNTSIPNVTEVKENMTFGSTLVTNPNGGFLACGPLYAYRCGHLHYTTGICSDVSPTFQVVNSIAPVQECSTQLDIVIVLDGSNSIYPWDSVTAFLNDLLERMDIGPKQTQVGIVQYGENVTHEFNLNKYSSTEEVLVAAKKIVQRGGRQTMTALGIDTARKEAFTEARGARRGVKKVMVIVTDGESHDNHRLKKVIQDCEDENIQRFSIAILGSYNRGNLSTEKFVEEIKSIASEPTEKHFFNVSDELALVTIVKTLGERIFALEATADQSAASFEMEMSQTGFSAHYSQDWVMLGAVGAYDWNGTVVMQKASQIIIPRNTTFNVESTKKNEPLASYLGYTVNSATASSGDVLYIAGQPRYNHTGQVIIYRMEDGNIKILQTLSGEQIGSYFGSILTTTDIDKDSNTDILLVGAPMYMGTEKEEQGKVYVYALNQTRFEYQMSLEPIKQTCCSSRQHNSCTTENKNEPCGARFGTAIAAVKDLNLDGFNDIVIGAPLEDDHGGAVYIYHGSGKTIRKEYAQRIPSGGDGKTLKFFGQSIHGEMDLNGDGLTDVTIGGLGGAALFWSRDVAVVKVTMNFEPNKVNIQKKNCHMEGKETVCINATVCFDVKLKSKEDTIYEADLQYRVTLDSLRQISRSFFSGTQERKVQRNITVRKSECTKHSFYMLDKHDFQDSVRITLDFNLTDPENGPVLDDSLPNSVHEYIPFAKDCGNKEKCISDLSLHVATTEKDLLIVRSQNDKFNVSLTVKNTKDSAYNTRTIVHYSPNLVFSGIEAIQKDSCESNHNITCKVGYPFLRRGEMVTFKILFQFNTSYLMENVTIYLSATSDSEEPPETLSDNVVNISIPVKYEVGLQFYSSASEYHISIAANETVPEVINSTEDIGNEINIFYLIRKSGSFPMPELKLSISFPNMTSNGYPVLYPTGLSSSENANCRPHIFEDPFSINSGKKMTTSTDHLKRGTILDCNTCKFATITCNLTSSDISQVNVSLILWKPTFIKSYFSSLNLTIRGELRSENASLVLSSSNQKRELAIQISKDGLPGRVPLWVILLSAFAGLLLLMLLILALWKIGFFKRPLKKKMEK tln1_mouse Talin-1 Tln1 Tln protein protein mouse Mus musculus Mouse MVALSLKISIGNVVKTMQFEPSTMVYDACRMIRERIPEALAGPPNDFGLFLSDDDPKKGIWLEAGKALDYYMLRNGDTMEYRKKQRPLKIRMLDGTVKTIMVDDSKTVTDMLMTICARIGITNHDEYSLVRELMEEKKDEGTGTLRKDKTLLRDEKKMEKLKQKLHTDDELNWLDHGRTLREQGVEEHETLLLRRKFFYSDQNVDSRDPVQLNLLYVQARDDILNGSHPVSFDKACEFAGFQCQIQFGPHNEQKHKAGFLDLKDFLPKEYVKQKGERKIFQAHKNCGQMSEIEAKVRYVKLARSLKTYGVSFFLVKEKMKGKNKLVPRLLGITKECVMRVDEKTKEVIQEWSLTNIKRWAASPKSFTLDFGDYQDGYYSVQTTEGEQIAQLIAGYIDIILKKKKSKDHFGLEGDEESTMLEDSVSPKKSTVLQQQYNRVGKVEHGSVALPAIMRSGASGPENFQVGSMPPAQQQITSGQMHRGHMPPLTSAQQALTGTINSSMQAVQAAQATLDDFETLPPLGQDAASKAWRKNKMDESKHEIHSQVDAITAGTASVVNLTAGDPAETDYTAVGCAVTTISSNLTEMSRGVKLLAALLEDEGGNGRPLLQAAKGLAGAVSELLRSAQPASAEPRQNLLQAAGNVGQASGELLQQIGESDTDPHFQDVLMQLAKAVASAAAALVLKAKSVAQRTEDSGLQTQVIAAATQCALSTSQLVACTKVVAPTISSPVCQEQLVEAGRLVAKAVEGCVSASQAATEDGQLLRGVGAAATAVTQALNELLQHVKAHATGAGPAGRYDQATDTILTVTENIFSSMGDAGEMVRQARILAQATSDLVNAIKADAEGESDLENSRKLLSAAKILADATAKMVEAAKGAAAHPDSEEQQQRLREAAEGLRMATNAAAQNAIKKKLVQRLEHAAKQAAASATQTIAAAQHAASAPKASAGPQPLLVQSCKAVAEQIPLLVQGVRGSQAQPDSPSAQLALIAASQSFLQPGGKMVAAAKASVPTIQDQASAMQLSQCAKNLGTALAELRTAAQKAQEACGPLEMDSALSVVQNLEKDLQEIKAAARDGKLKPLPGETMEKCTQDLGNSTKAVSSAIAKLLGEIAQGNENYAGIAARDVAGGLRSLAQAARGVAALTSDPAVQAIVLDTASDVLDKASSLIEEAKKASGHPGDPESQQRLAQVAKAVTQALNRCVSCLPGQRDVDNALRAVGDASKRLLSDSLPPSTGTFQEAQSRLNEAAAGLNQAATELVQASRGTPQDLARASGRFGQDFSTFLEAGVEMAGQAPSQEDRAQVVSNLKGISMSSSKLLLAAKALSTDPASPNLKSQLAAAARAVTDSINQLITMCTQQAPGQKECDNALRQLETVRELLENPVQPINDMSYFGCLDSVMENSKVLGEAMTGISQNAKNGNLPEFGDAIATASKALCGFTEAAAQAAYLVGVSDPNSQAGQQGLVEPTQFARANQAIQMACQSLGEPGCTQAQVLSAATIVAKHTSALCNSCRLASARTANPTAKRQFVQSAKEVANSTANLVKTIKALDGDFTEENRAQCRAATAPLLEAVDNLSAFASNPEFSSVPAQISPEGRAAMEPIVISAKTMLESAGGLIQTARALAVNPRDPPRWSVLAGHSRTVSDSIKKLITSMRDKAPGQLECETAIAALNSCLRDLDQASLAAVSQQLAPREGISQEALHTQMLTAVQEISHLIEPLASAARAEASQLGHKVSQMAQYFEPLTLAAVGAASKTLSHPQQMALLDQTKTLAESALQLLYTAKEAGGNPKQAAHTQEALEEAVQMMTEAVEDLTTTLNEAASAAGVVGGMVDSITQAINQLDEGPMGDPEGSFVDYQTTMVRTAKAIAVTVQEMVTKSNTSPEELGPLANQLTSDYGRLASQAKPAAVAAENEEIGAHIKHRVQELGHGCSALVTKAGALQCSPSDVYTKKELIECARRVSEKVSHVLAALQAGNRGTQACITAASAVSGIIADLDTTIMFATAGTLNREGAETFADHREGILKTAKVLVEDTKVLVQNAAGSQEKLAQAAQSSVATITRLADVVKLGAASLGAEDPETQVVLINAVKDVAKALGDLISATKAAAGKVGDDPAVWQLKNSAKVMVTNVTSLLKTVKAVEDEATKGTRALEATTEHIRQELAVFCSPEPPAKTSTPEDFIRMTKGITMATAKAVAAGNSCRQEDVIATANLSRRAIADMLRACKEAAFHPEVAPDVRLRALHYGRECANGYLELLDHVLLTLQKPNPDLKQQLTGHSKRVAGSVTELIQAAEAMKGTEWVDPEDPTVIAENELLGAAAAIEAAAKKLEQLKPRAKPKEADESLNFEEQILEAAKSIAAATSALVKAASAAQRELVAQGKVGAIPANALDDGQWSQGLISAARMVAAATNNLCEAANAAVQGHASQEKLISSAKQVAASTAQLLVACKVKADQDSEAMKRLQAAGNAVKRASDNLVKAAQKAAAFEDQENETVVVKEKMVGGIAQIIAAQEEMLRKERELEEARKKLAQIRQQQYKFLPSELRDEH itb1_mouse Integrin beta-1 Fibronectin receptor subunit beta VLA-4 subunit beta Itgb1 protein protein mouse Mus musculus Mouse MNLQLVSWIGLISLICSVFGQTDKNRCLKANAKSCGECIQAGPNCGWCTNTTFLQEGMPTSARCDDLEALKKKGCQPSDIENPRGSQTIKKNKNVTNRSKGMAEKLRPEDITQIQPQQLLLKLRSGEPQKFTLKFKRAEDYPIDLYYLMDLSYSMKDDLENVKSLGTDLMNEMRRITSDFRIGFGSFVEKTVMPYISTTPAKLRNPCTSEQNCTSPFSYKNVLSLTDRGEFFNELVGQQRISGNLDSPEGGFDAIMQVAVCGSLIGWRNVTRLLVFSTDAGFHFAGDGKLGGIVLPNDGQCHLENNVYTMSHYYDYPSIAHLVQKLSENNIQTIFAVTEEFQPVYKELKNLIPKSAVGTLSGNSSNVIQLIIDAYNSLSSEVILENSKLPDGVTINYKSYCKNGVNGTGENGRKCSNISIGDEVQFEISITANKCPNKESETIKIKPLGFTEEVEVVLQFICKCNCQSHGIPASPKCHEGNGTFECGACRCNEGRVGRHCECSTDEVNSEDMDAYCRKENSSEICSNNGECVCGQCVCRKRDNTNEIYSGKFCECDNFNCDRSNGLICGGNGVCRCRVCECYPNYTGSACDCSLDTGPCLASNGQICNGRGICECGACKCTDPKFQGPTCETCQTCLGVCAEHKECVQCRAFNKGEKKDTCAQECSHFNLTKVESREKLPQPVQVDPVTHCKEKDIDDCWFYFTYSVNGNNEAIVHVVETPDCPTGPDIIPIVAGVVAGIVLIGLALLLIWKLLMIIHDRREFAKFEKEKMNAKWDTGENPIYKSAVTTVVNPKYEGK ferm2_mouse Fermitin family homolog 2 Fermt2 Plekhc1 Kindlin-2 Pleckstrin homology domain-containing family C member 1 protein protein mouse Mus musculus Mouse MALDGIRMPDGCYADGTWELSVHVTDLNRDVTLRVTGEVHIGGVMLKLVEKLDVKKDWSDHALWWEKKRTWLLKTHWTLDKCGIQADAKLQFTPQHKLLRLQLPNMKYVKVKVNFSDRVFKAVSDICKTFNIRHPEELSLLKKPRDPTKKKKKKLDDQSEDEALELEGPLIMPGSGSIYSSPGLYSKTMTPTYDAHDGSPLSPTSAWFGDSALSEGNPGILAVSQPVTSPEILAKMFKPQALLDKAKTNQGWLDSSRSLMEQDVKENEALLLRFKYYSFFDLNPKYDAIRINQLYEQAKWALLLEEIECTEEEMMMFAALQYHINKLSIMTSENHLNNSDKEVDEVDAALSDLEITLEGGKTSTILGDITSIPELADYIKVFKPKKLTLKGYKQYWCTFKDTSISCYKSREESSGTPAHQLNLRGCEVTPDVNISGQKFNIKLLIPVAEGMNEIWLRCDNEKQYAHWMAACRLASKGKTMADSSYNLEVQNILSFLKMQHLNPDPQLIPDQITTDVNPECLVSPRYLKKYKSKQITARILEAHQNVAQMSLIEAKMRFIQAWQSLPEFGITHFIARFQGGKREELIGIAYNRLIRMDASTGDAIKTWRFSNMKQWNVNWEIKMVTVEFADEVRLSFICTEVDCKVVHEFIGGYIFLSTRAKDQNESLDEEMFYKLTSGWV itga2-itgb1-2 1270157 n/a 1270167 unspecified role unspecified role prey prey n/a 1270168 unspecified role unspecified role bait bait n/a 1270169 unspecified role unspecified role prey prey association association false false false Fig 3c, 3d Using PC3 cells grown on plastic, SHARPIN co-precipitated strongly with alpha2-beta1-integrin and, to a lesser extent, with alpha5-beta1-integrin. SHARPIN coimmunoprecipitated more strongly with alpha-integrins from cells grown in suspension, rather than cells grown adhering to their substrate collagens. itga5-itgb1-1 1270157 n/a 1270167 unspecified role unspecified role prey prey n/a 1270170 unspecified role unspecified role bait bait n/a 1270169 unspecified role unspecified role prey prey association association false false false Fig 3c Using PC3 cells grown on plastic, SHARPIN co-precipitated strongly with alpha2-beta1-integrin and, to a lesser extent, with alpha5-beta1-integrin. il1a-sharpin 1270157 n/a 1270169 unspecified role unspecified role prey prey n/a 1270171 unspecified role unspecified role bait bait physical association physical association false false false fig 3e SHARPIN coimmunoprecipitated more strongly with alpha-integrins from cells grown in suspension, rather than cells grown adhering to their substrate collagens. itga2-sharpin 1270158 n/a 1270172 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 false n/a 1270168 unspecified role unspecified role bait bait conserved_membrane_proximal_segment sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false Fig 2b itga1-sharpin-1 1270158 n/a 1270172 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 false n/a 1270173 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false c-terminus sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false Fig 2b SHARPIN associates with the cytoplasmic domain of integrin alpha-subunits. itga2-sharpin-2 1270159 n/a 1270169 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 false ecoli Escherichia coli (strain K12) n/a 1270168 unspecified role unspecified role bait bait c_terminus sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false Fig 2d, 2f. itgb1-itgb1-3 1270159 n/a 1270174 unspecified role unspecified role prey prey head_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1270175 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false cytoplasmic domain sufficient to bind sufficient binding region sufficient to bind less-than less-than certain certain sequence position certain false direct interaction direct interaction false false false Fig 2d itga2-sharpin-5 1270160 n/a 1270168 unspecified role unspecified role bait bait c_terminus sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ilr2alpha_tac_subunit_ec_and tm_domains fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1270169 unspecified role unspecified role prey prey 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 false physical association physical association false false false Fig 2g The integrin alpha-subunit binding sequence interacts with SHARPIN even when inserted into an irrelevant transmembrane protein. itga5-sharpin 1270160 n/a 1270170 unspecified role unspecified role bait bait c_terminus sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ilr2alpha_tac_subunit_ec_and tm_domains fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1270169 unspecified role unspecified role prey prey 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 false physical association physical association false false false Fig 2g The integrin alpha-subunit binding sequence interacts with SHARPIN even when inserted into an irrelevant transmembrane protein. itgb1-tln1 1270161 n/a 1270174 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1270167 unspecified role unspecified role fluorescence donor fluorescence donor 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 false proximity proximity close-contact colocalization neighbourhood interaction false false false fig 4b There was a significant reduction in the interaction between beta1–GFP and mCherry–talin and beta1–GFP and mCherry–kindlin-2 in cells positive for MYC–SHARPIN. itgb1-tln1-1 1270161 n/a 1270167 unspecified role unspecified role fluorescence donor fluorescence donor 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 false n/a 1270176 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false proximity proximity close-contact colocalization neighbourhood interaction false false false fig 4c There was a significant reduction in the interaction between beta1–GFP and mCherry–talin and beta1–GFP and mCherry–kindlin-2 in cells positive for MYC–SHARPIN. itga2-sharpin-1 1270162 n/a 1270168 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false conserved_membrane_proximal_segment sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1270169 unspecified role unspecified role prey prey physical association physical association false false false Fig 2c itga2-sharpin-4 1270163 n/a 1270169 unspecified role unspecified role neutral component neutral component 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 false n/a 1270168 unspecified role unspecified role neutral component neutral component conserved_membrane_proximal_segment sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false edans_tag fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P17301:p.Trp1154_Phe1159delinsAlaAlaAlaAlaAlaAla mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false 1270163 Fig 2e SHARPIN binding is mainly mediated by residues WK, LG and FF. itgb1-sharpin-3 1270164 n/a 1270169 unspecified role unspecified role neutral component neutral component n/a 1270167 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Fig 3 SHARPIN co-localizes with inactive beta1-integrins in cells, but not with active beta1-integrins. itga5-sharpin 1270165 n/a 1270169 unspecified role unspecified role prey prey n/a 1270170 unspecified role unspecified role bait bait 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 false P08648:p.Leu1023_Gly1024delinsAlaAla mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false fig 3f itga5-sharpin-2 1270166 n/a 1270170 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept region mcherry mcherry fluorescent protein tag mcherry undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1270169 unspecified role unspecified role fluorescence donor fluorescence donor 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 false proximity proximity close-contact colocalization neighbourhood interaction false false false Fig 4a