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/ wegener-2007-1 Structural basis of integrin activation by talin. in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Wegener KL., Partridge AW., Han J., Pickford AR., Liddington RC., Ginsberg MH., Campbell ID. 1H-15N-HSQC titrations were performed. The cys336ser talin-1 mutant was used in all studies. The fact mouse talin-1 not -2 was used was found by looking at the reference de Pereda et al. (2005) as only talin-1 has a Cys at position 336. iain.campbell@bioch.ox.ac.uk,mhginsberg@ucsd.edu Cell (0092-8674) 2007 Only protein-protein interactions 7 imex curation wegener-2007-2 Structural basis of integrin activation by talin. in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Wegener KL., Partridge AW., Han J., Pickford AR., Liddington RC., Ginsberg MH., Campbell ID. 1H-15N-HSQC titrations were performed. The cys336ser talin-1 mutant was used in all studies. The fact mouse talin-1 not -2 was used was found by looking at the reference de Pereda et al. (2005) as only talin-1 has a Cys at position 336. iain.campbell@bioch.ox.ac.uk,mhginsberg@ucsd.edu Cell (0092-8674) 2007 Only protein-protein interactions 7 imex curation wegener-2007-3 Structural basis of integrin activation by talin. in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Wegener KL., Partridge AW., Han J., Pickford AR., Liddington RC., Ginsberg MH., Campbell ID. 1H-15N-HSQC titrations were performed. The cys336ser talin-1 mutant was used in all studies. The fact mouse talin-1 not -2 was used was found by looking at the reference de Pereda et al. (2005) as only talin-1 has a Cys at position 336. iain.campbell@bioch.ox.ac.uk,mhginsberg@ucsd.edu Cell (0092-8674) 2007 Only protein-protein interactions 7 imex curation tln1_mouse Talin-1 Tln1 Tln protein protein mouse Mus musculus Mouse MVALSLKISIGNVVKTMQFEPSTMVYDACRMIRERIPEALAGPPNDFGLFLSDDDPKKGIWLEAGKALDYYMLRNGDTMEYRKKQRPLKIRMLDGTVKTIMVDDSKTVTDMLMTICARIGITNHDEYSLVRELMEEKKDEGTGTLRKDKTLLRDEKKMEKLKQKLHTDDELNWLDHGRTLREQGVEEHETLLLRRKFFYSDQNVDSRDPVQLNLLYVQARDDILNGSHPVSFDKACEFAGFQCQIQFGPHNEQKHKAGFLDLKDFLPKEYVKQKGERKIFQAHKNCGQMSEIEAKVRYVKLARSLKTYGVSFFLVKEKMKGKNKLVPRLLGITKECVMRVDEKTKEVIQEWSLTNIKRWAASPKSFTLDFGDYQDGYYSVQTTEGEQIAQLIAGYIDIILKKKKSKDHFGLEGDEESTMLEDSVSPKKSTVLQQQYNRVGKVEHGSVALPAIMRSGASGPENFQVGSMPPAQQQITSGQMHRGHMPPLTSAQQALTGTINSSMQAVQAAQATLDDFETLPPLGQDAASKAWRKNKMDESKHEIHSQVDAITAGTASVVNLTAGDPAETDYTAVGCAVTTISSNLTEMSRGVKLLAALLEDEGGNGRPLLQAAKGLAGAVSELLRSAQPASAEPRQNLLQAAGNVGQASGELLQQIGESDTDPHFQDVLMQLAKAVASAAAALVLKAKSVAQRTEDSGLQTQVIAAATQCALSTSQLVACTKVVAPTISSPVCQEQLVEAGRLVAKAVEGCVSASQAATEDGQLLRGVGAAATAVTQALNELLQHVKAHATGAGPAGRYDQATDTILTVTENIFSSMGDAGEMVRQARILAQATSDLVNAIKADAEGESDLENSRKLLSAAKILADATAKMVEAAKGAAAHPDSEEQQQRLREAAEGLRMATNAAAQNAIKKKLVQRLEHAAKQAAASATQTIAAAQHAASAPKASAGPQPLLVQSCKAVAEQIPLLVQGVRGSQAQPDSPSAQLALIAASQSFLQPGGKMVAAAKASVPTIQDQASAMQLSQCAKNLGTALAELRTAAQKAQEACGPLEMDSALSVVQNLEKDLQEIKAAARDGKLKPLPGETMEKCTQDLGNSTKAVSSAIAKLLGEIAQGNENYAGIAARDVAGGLRSLAQAARGVAALTSDPAVQAIVLDTASDVLDKASSLIEEAKKASGHPGDPESQQRLAQVAKAVTQALNRCVSCLPGQRDVDNALRAVGDASKRLLSDSLPPSTGTFQEAQSRLNEAAAGLNQAATELVQASRGTPQDLARASGRFGQDFSTFLEAGVEMAGQAPSQEDRAQVVSNLKGISMSSSKLLLAAKALSTDPASPNLKSQLAAAARAVTDSINQLITMCTQQAPGQKECDNALRQLETVRELLENPVQPINDMSYFGCLDSVMENSKVLGEAMTGISQNAKNGNLPEFGDAIATASKALCGFTEAAAQAAYLVGVSDPNSQAGQQGLVEPTQFARANQAIQMACQSLGEPGCTQAQVLSAATIVAKHTSALCNSCRLASARTANPTAKRQFVQSAKEVANSTANLVKTIKALDGDFTEENRAQCRAATAPLLEAVDNLSAFASNPEFSSVPAQISPEGRAAMEPIVISAKTMLESAGGLIQTARALAVNPRDPPRWSVLAGHSRTVSDSIKKLITSMRDKAPGQLECETAIAALNSCLRDLDQASLAAVSQQLAPREGISQEALHTQMLTAVQEISHLIEPLASAARAEASQLGHKVSQMAQYFEPLTLAAVGAASKTLSHPQQMALLDQTKTLAESALQLLYTAKEAGGNPKQAAHTQEALEEAVQMMTEAVEDLTTTLNEAASAAGVVGGMVDSITQAINQLDEGPMGDPEGSFVDYQTTMVRTAKAIAVTVQEMVTKSNTSPEELGPLANQLTSDYGRLASQAKPAAVAAENEEIGAHIKHRVQELGHGCSALVTKAGALQCSPSDVYTKKELIECARRVSEKVSHVLAALQAGNRGTQACITAASAVSGIIADLDTTIMFATAGTLNREGAETFADHREGILKTAKVLVEDTKVLVQNAAGSQEKLAQAAQSSVATITRLADVVKLGAASLGAEDPETQVVLINAVKDVAKALGDLISATKAAAGKVGDDPAVWQLKNSAKVMVTNVTSLLKTVKAVEDEATKGTRALEATTEHIRQELAVFCSPEPPAKTSTPEDFIRMTKGITMATAKAVAAGNSCRQEDVIATANLSRRAIADMLRACKEAAFHPEVAPDVRLRALHYGRECANGYLELLDHVLLTLQKPNPDLKQQLTGHSKRVAGSVTELIQAAEAMKGTEWVDPEDPTVIAENELLGAAAAIEAAAKKLEQLKPRAKPKEADESLNFEEQILEAAKSIAAATSALVKAASAAQRELVAQGKVGAIPANALDDGQWSQGLISAARMVAAATNNLCEAANAAVQGHASQEKLISSAKQVAASTAQLLVACKVKADQDSEAMKRLQAAGNAVKRASDNLVKAAQKAAAFEDQENETVVVKEKMVGGIAQIIAAQEEMLRKERELEEARKKLAQIRQQQYKFLPSELRDEH itb3_human Integrin beta-3 Platelet membrane glycoprotein IIIa ITGB3 GP3A protein protein human Homo sapiens Human MRARPRPRPLWATVLALGALAGVGVGGPNICTTRGVSSCQQCLAVSPMCAWCSDEALPLGSPRCDLKENLLKDNCAPESIEFPVSEARVLEDRPLSDKGSGDSSQVTQVSPQRIALRLRPDDSKNFSIQVRQVEDYPVDIYYLMDLSYSMKDDLWSIQNLGTKLATQMRKLTSNLRIGFGAFVDKPVSPYMYISPPEALENPCYDMKTTCLPMFGYKHVLTLTDQVTRFNEEVKKQSVSRNRDAPEGGFDAIMQATVCDEKIGWRNDASHLLVFTTDAKTHIALDGRLAGIVQPNDGQCHVGSDNHYSASTTMDYPSLGLMTEKLSQKNINLIFAVTENVVNLYQNYSELIPGTTVGVLSMDSSNVLQLIVDAYGKIRSKVELEVRDLPEELSLSFNATCLNNEVIPGLKSCMGLKIGDTVSFSIEAKVRGCPQEKEKSFTIKPVGFKDSLIVQVTFDCDCACQAQAEPNSHRCNNGNGTFECGVCRCGPGWLGSQCECSEEDYRPSQQDECSPREGQPVCSQRGECLCGQCVCHSSDFGKITGKYCECDDFSCVRYKGEMCSGHGQCSCGDCLCDSDWTGYYCNCTTRTDTCMSSNGLLCSGRGKCECGSCVCIQPGSYGDTCEKCPTCPDACTFKKECVECKKFDRGALHDENTCNRYCRDEIESVKELKDTGKDAVNCTYKNEDDCVVRFQYYEDSSGKSILYVVEEPECPKGPDILVVLLSVMGAILLIGLAALLIWKLLITIHDRKEFAKFEEERARAKWDTANNPLYKEATSTFTNITYRGT itgb3_pipkig_hum_pep Synthetic chimeric peptide of human integrin beta3 membrane proximal peptide (742-765) and human PIPKIgamma C-terminus (648-657) peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) KLLITIHDRKEFAKFEEERARAKWVYSPLHYSAR An amino acid sequence which is not represented in UniProt The residue in the peptide corresponding to residue 657 of PIPKIgamma is Arg in this peptide but Glu in the Uniprotkb O60331 sequence. tln1-itgb3 NMR titration shows talin F3 domain interacts with membrane proximal peptide of the integrin beta3 tail 576250 N/A 576253 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) N/A 576254 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false 1 A, B, S1 It is not clear whether the Integrin beta-3 peptide was chemically synthesized or produced as a GST-beta3 recombinant fusion protein and liberated by thrombin digetion. The coordinates given in the paper for the integrin beta3 tail (peptides) are different to those in the UniProt entry. tln1-itgb3-1 NMR titration shows talin F3 domain interacts with membrane distal peptide of the integrin beta3 tail 576250 N/A 576254 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 576253 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false 1 A, C, S1 It is not clear whether the Integrin beta-3 peptide was chemically synthesized or produced as a GST-beta3 recombinant fusion protein and liberated by thrombin digetion. The coordinates given in the paper for the Integrin beta-3 tail (peptides) are different to those in the UniProt entry. tln1-itgb3-2 NMR titration shows talin F3 domain interacts with the integrin beta3 tail 576250 N/A 576253 unspecified role unspecified role neutral component neutral component P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false region 15n label 15n label 15N N15 15N N15 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 ecobb Escherichia coli (strain B / BL21) N/A 576254 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false 1 A, S1 It is not clear whether the Integrin beta-3 peptide was chemically synthesized or produced as a GST-beta3 recombinant fusion protein and liberated by thrombin digetion. The coordinates given in the paper for the integrin beta3 tail (peptides) are different to those in the UniProt entry. tln1-itgb3_pipkig_hu NMR titration shows talin F3 domain interacts with the chimeric integrin beta3 tail-PIPKIgamma C-terminus peptide 576250 N/A 576255 unspecified role unspecified role neutral component neutral component phe12ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false phe15ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain false Stoichiometry: 1.0 N/A 576253 unspecified role unspecified role neutral component neutral component P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false region 15n label 15n label 15N N15 15N N15 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 ecobb Escherichia coli (strain B / BL21) Stoichiometry: 1.0 direct interaction direct interaction false false false 1 A, 4, S1 Binding to the MP region of the chimeric peptide by talin is reduced by phe12ala and abolished by phe15ala in the chimeric peptide. The coordinates given in the paper for the integrin beta3 tail (peptides) are different to those in the UniProt entry. tln1-itgb3_pipkig_hu NMR titration shows talin F3 domain interacts with the chimeric integrin beta3 tail-PIPKIgamma C-terminus peptide 576250 N/A 576253 unspecified role unspecified role neutral component neutral component region 15n label 15n label 15N N15 15N N15 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 P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false P26039:p.Leu325Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false P26039:p.Gln381Val mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) Stoichiometry: 1.0 N/A 576255 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 direct interaction direct interaction false false false 1 A, 4, S1 Binding to the MP region of the chimeric peptide by talin is reduced by glu381val and abolished by leu325arg in talin F3. The coordinates given in the paper for the integrin beta3 tail (peptides) are different to those in the UniProt entry. tln1-itgb3_pipkig_hu NMR structural study shows talin F3 interacts with chimeric integrin beta3 tail-PIPKIgamma C-terminus peptide 576251 N/A 576255 unspecified role unspecified role neutral component neutral component N/A 576253 unspecified role unspecified role neutral component neutral component region 15n label 15n label 15N N15 15N N15 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 P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false region 2h label 2h label deuterium D2 2H2 2H2 deuterium D2 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction false false false Table S1 tln1-itgb3_pipkig_hu ITC shows talin F3 domain interacts with the chimeric integrin beta3 tail-PIPKIgamma C-terminus peptide 576252 N/A 576253 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain false P26039:p.Leu325Arg mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P26039:p.Trp359Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ecobb Escherichia coli (strain B / BL21) Stoichiometry: 1.0 N/A 576255 unspecified role unspecified role neutral component neutral component Stoichiometry: 1.0 direct interaction direct interaction false false false Table S2, S5 Talin F3 trp359ala Kd = 1.06 E-4 M; Talin F3 leu325arg Kd= 2.12 E-7 M