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/ Structural basis of integrin activation by talin. Structural basis of integrin activation by talin. Cell (0092-8674) 2007 imex curation 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 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 7 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 Accepted 2007-FEB-8 by DAVET. 02-MAR-2006: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 7 imex curation Structural basis of integrin activation by talin. Structural basis of integrin activation by talin. Cell (0092-8674) 2007 imex curation 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 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 7 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 Accepted 2007-FEB-8 by DAVET. 02-MAR-2006: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 7 imex curation Structural basis of integrin activation by talin. Structural basis of integrin activation by talin. Cell (0092-8674) 2007 imex curation 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 02-MAR-2006: Contacted by JYOTI. Only protein-protein interactions 7 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 Accepted 2007-FEB-8 by DAVET. 02-MAR-2006: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 7 imex curation tln1_mouse Talin-1 Tln1 Tln protein protein mouse Mus musculus Mouse MVALSLKISIGNVVKTMQFEPSTMVYDACRMIRERIPEALAGPPNDFGLFLSDDDPKKGIWLEAGKALDYYMLRNGDTMEYRKKQRPLKIRMLDGTVKTIMVDDSKTVTDMLMTICARIGITNHDEYSLVRELMEEKKDEGTGTLRKDKTLLRDEKKMEKLKQKLHTDDELNWLDHGRTLREQGVEEHETLLLRRKFFYSDQNVDSRDPVQLNLLYVQARDDILNGSHPVSFDKACEFAGFQCQIQFGPHNEQKHKAGFLDLKDFLPKEYVKQKGERKIFQAHKNCGQMSEIEAKVRYVKLARSLKTYGVSFFLVKEKMKGKNKLVPRLLGITKECVMRVDEKTKEVIQEWSLTNIKRWAASPKSFTLDFGDYQDGYYSVQTTEGEQIAQLIAGYIDIILKKKKSKDHFGLEGDEESTMLEDSVSPKKSTVLQQQYNRVGKVEHGSVALPAIMRSGASGPENFQVGSMPPAQQQITSGQMHRGHMPPLTSAQQALTGTINSSMQAVQAAQATLDDFETLPPLGQDAASKAWRKNKMDESKHEIHSQVDAITAGTASVVNLTAGDPAETDYTAVGCAVTTISSNLTEMSRGVKLLAALLEDEGGNGRPLLQAAKGLAGAVSELLRSAQPASAEPRQNLLQAAGNVGQASGELLQQIGESDTDPHFQDVLMQLAKAVASAAAALVLKAKSVAQRTEDSGLQTQVIAAATQCALSTSQLVACTKVVAPTISSPVCQEQLVEAGRLVAKAVEGCVSASQAATEDGQLLRGVGAAATAVTQALNELLQHVKAHATGAGPAGRYDQATDTILTVTENIFSSMGDAGEMVRQARILAQATSDLVNAIKADAEGESDLENSRKLLSAAKILADATAKMVEAAKGAAAHPDSEEQQQRLREAAEGLRMATNAAAQNAIKKKLVQRLEHAAKQAAASATQTIAAAQHAASAPKASAGPQPLLVQSCKAVAEQIPLLVQGVRGSQAQPDSPSAQLALIAASQSFLQPGGKMVAAAKASVPTIQDQASAMQLSQCAKNLGTALAELRTAAQKAQEACGPLEMDSALSVVQNLEKDLQEIKAAARDGKLKPLPGETMEKCTQDLGNSTKAVSSAIAKLLGEIAQGNENYAGIAARDVAGGLRSLAQAARGVAALTSDPAVQAIVLDTASDVLDKASSLIEEAKKASGHPGDPESQQRLAQVAKAVTQALNRCVSCLPGQRDVDNALRAVGDASKRLLSDSLPPSTGTFQEAQSRLNEAAAGLNQAATELVQASRGTPQDLARASGRFGQDFSTFLEAGVEMAGQAPSQEDRAQVVSNLKGISMSSSKLLLAAKALSTDPASPNLKSQLAAAARAVTDSINQLITMCTQQAPGQKECDNALRQLETVRELLENPVQPINDMSYFGCLDSVMENSKVLGEAMTGISQNAKNGNLPEFGDAIATASKALCGFTEAAAQAAYLVGVSDPNSQAGQQGLVEPTQFARANQAIQMACQSLGEPGCTQAQVLSAATIVAKHTSALCNSCRLASARTANPTAKRQFVQSAKEVANSTANLVKTIKALDGDFTEENRAQCRAATAPLLEAVDNLSAFASNPEFSSVPAQISPEGRAAMEPIVISAKTMLESAGGLIQTARALAVNPRDPPRWSVLAGHSRTVSDSIKKLITSMRDKAPGQLECETAIAALNSCLRDLDQASLAAVSQQLAPREGISQEALHTQMLTAVQEISHLIEPLASAARAEASQLGHKVSQMAQYFEPLTLAAVGAASKTLSHPQQMALLDQTKTLAESALQLLYTAKEAGGNPKQAAHTQEALEEAVQMMTEAVEDLTTTLNEAASAAGVVGGMVDSITQAINQLDEGPMGDPEGSFVDYQTTMVRTAKAIAVTVQEMVTKSNTSPEELGPLANQLTSDYGRLASQAKPAAVAAENEEIGAHIKHRVQELGHGCSALVTKAGALQCSPSDVYTKKELIECARRVSEKVSHVLAALQAGNRGTQACITAASAVSGIIADLDTTIMFATAGTLNREGAETFADHREGILKTAKVLVEDTKVLVQNAAGSQEKLAQAAQSSVATITRLADVVKLGAASLGAEDPETQVVLINAVKDVAKALGDLISATKAAAGKVGDDPAVWQLKNSAKVMVTNVTSLLKTVKAVEDEATKGTRALEATTEHIRQELAVFCSPEPPAKTSTPEDFIRMTKGITMATAKAVAAGNSCRQEDVIATANLSRRAIADMLRACKEAAFHPEVAPDVRLRALHYGRECANGYLELLDHVLLTLQKPNPDLKQQLTGHSKRVAGSVTELIQAAEAMKGTEWVDPEDPTVIAENELLGAAAAIEAAAKKLEQLKPRAKPKEADESLNFEEQILEAAKSIAAATSALVKAASAAQRELVAQGKVGAIPANALDDGQWSQGLISAARMVAAATNNLCEAANAAVQGHASQEKLISSAKQVAASTAQLLVACKVKADQDSEAMKRLQAAGNAVKRASDNLVKAAQKAAAFEDQENETVVVKEKMVGGIAQIIAAQEEMLRKERELEEARKKLAQIRQQQYKFLPSELRDEH DIP protein P26039 original sequence version: 109 DIP protein P26039 original sequence version: 114 DIP protein P26039 original sequence version: 151 DIP protein P26039 original sequence version: 165 DIP protein P26039 original sequence version: 136 78832388E2392B8E itb3_human Integrin beta-3 Platelet membrane glycoprotein IIIa ITGB3 GP3A protein protein human Homo sapiens Human MRARPRPRPLWATVLALGALAGVGVGGPNICTTRGVSSCQQCLAVSPMCAWCSDEALPLGSPRCDLKENLLKDNCAPESIEFPVSEARVLEDRPLSDKGSGDSSQVTQVSPQRIALRLRPDDSKNFSIQVRQVEDYPVDIYYLMDLSYSMKDDLWSIQNLGTKLATQMRKLTSNLRIGFGAFVDKPVSPYMYISPPEALENPCYDMKTTCLPMFGYKHVLTLTDQVTRFNEEVKKQSVSRNRDAPEGGFDAIMQATVCDEKIGWRNDASHLLVFTTDAKTHIALDGRLAGIVQPNDGQCHVGSDNHYSASTTMDYPSLGLMTEKLSQKNINLIFAVTENVVNLYQNYSELIPGTTVGVLSMDSSNVLQLIVDAYGKIRSKVELEVRDLPEELSLSFNATCLNNEVIPGLKSCMGLKIGDTVSFSIEAKVRGCPQEKEKSFTIKPVGFKDSLIVQVTFDCDCACQAQAEPNSHRCNNGNGTFECGVCRCGPGWLGSQCECSEEDYRPSQQDECSPREGQPVCSQRGECLCGQCVCHSSDFGKITGKYCECDDFSCVRYKGEMCSGHGQCSCGDCLCDSDWTGYYCNCTTRTDTCMSSNGLLCSGRGKCECGSCVCIQPGSYGDTCEKCPTCPDACTFKKECVECKKFDRGALHDENTCNRYCRDEIESVKELKDTGKDAVNCTYKNEDDCVVRFQYYEDSSGKSILYVVEEPECPKGPDILVVLLSVMGAILLIGLAALLIWKLLITIHDRKEFAKFEEERARAKWDTANNPLYKEATSTFTNITYRGT DIP protein P05106 original sequence version: 202 DIP protein P05106-1 original sequence version: 158 DIP protein P05106-1 original sequence version: 164 DIP protein P05106 original sequence version: 200 DIP protein P05106-1 original sequence version: 213 DIP protein P05106-1 original sequence version: 175 DIP protein P05106-1 original sequence version: 192 DIP protein P05106-1 original sequence version: 171 F246623608E05F9E 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. 754A0CFCF69549F8 tln1-itgb3 1 4 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 P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain C S Wed May 02 19:15:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys336ser' Tue Jun 30 19:20:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys336Ser' region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) 5 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 direct interaction direct interaction 1 A, B, S1 Weighted shift maps (see Experimental Procedures) of induced chemical shifts seen in 1H-15N-HSQC spectra when the (B) MP peptide and (C) MD peptides are added to the talin F3 subdomain. 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 1 5 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 4 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 P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:37:22 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys336ser' Tue Jun 30 18:04:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys336Ser' region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction 1 A, C, S1 Weighted shift maps (see Experimental Procedures) of induced chemical shifts seen in 1H-15N-HSQC spectra when the (B) MP peptide and (C) MD peptides are added to the talin F3 subdomain. 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 1 4 unspecified role unspecified role neutral component neutral component P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain C S Wed May 02 17:31:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys336ser' Tue Jun 30 19:33:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys336Ser' region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 5 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 direct interaction direct interaction 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. NMR studies of the interaction with full-length b3 cytoplasmic tails were more challenging. The off rates of the complex correspond to an "intermediate exchange" regime, with peaks broadening and disappearing rather than changing position. It was possible, however, to define perturbed residues by plotting the percentage decrease in peak height upon addition of the 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 1 6 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 F A phe15ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing alanine scanning alanine scanning certain certain sequence position certain certain certain sequence position certain F A 4 unspecified role unspecified role neutral component neutral component P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain C S Wed May 02 16:59:49 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys336ser' Tue Jun 30 18:21:42 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys336Ser' region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction 1 A, 4, S1 a synthetic chimeric peptide comprised of b3-integrin tail residues on the N-terminal side of the critical tryptophan and PIPKIg residues on the C-terminal side would yield a peptide ligand with higher affinity and better solubility that would be suitable for high-resolution studies of the F3-MP interaction. The sequence of the designed chimeric peptide is shown in Figure 1A. NMR experiments showed that this chimeric peptide does indeed bind to talin and forms a tight, highly soluble 1:1 complex. 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 3 6 unspecified role unspecified role neutral component neutral component 4 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain C S Wed May 02 19:16:02 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys336ser' Tue Jun 30 17:58:51 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys336Ser' region 2h label 2h label deuterium D2 2H2 2H2 deuterium D2 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Table S1 The Structure of the Talin F3/Peptide Complex Reveals a Binding Interface with the MP Site The structure of the complex was calculated from NMR data using a total of 2187 experimental restraints, 138 of which were unambiguous intermolecular nuclear Overhauser effects tln1-itgb3_pipkig_hu 1 4 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain C S Wed May 02 19:54:57 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys336ser' Tue Jun 30 19:58:25 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys336Ser' 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 L R Wed May 02 19:02:53 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu325arg' Tue Jun 30 17:09:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu325Arg' 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 Q V Wed May 02 16:16:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln381val' Tue Jun 30 18:34:21 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln381Val' ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role neutral component neutral component direct interaction direct interaction 1 A, 4, S1 a synthetic chimeric peptide comprised of b3-integrin tail residues on the N-terminal side of the critical tryptophan and PIPKIg residues on the C-terminal side would yield a peptide ligand with higher affinity and better solubility that would be suitable for high-resolution studies of the F3-MP interaction. The sequence of the designed chimeric peptide is shown in Figure 1A. NMR experiments showed that this chimeric peptide does indeed bind to talin and forms a tight, highly soluble 1:1 complex. 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 2 4 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 P26039:p.Cys336Ser mutation mutation certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:23:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys336ser' Tue Jun 30 19:32:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys336Ser' 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 L R Wed May 02 18:53:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu325arg' Tue Jun 30 18:58:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu325Arg' region 15n label 15n label 15N N15 15N N15 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P26039:p.Trp359Ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain W A Wed May 02 19:08:43 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'trp359ala' Tue Jun 30 17:27:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Trp359Ala' ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role neutral component neutral component direct interaction direct interaction Table S2, S5 the talin (W359A) mutant, which inhibits binding of the MD site of the b3 tail, reduced the affinity 1000- fold, Talin F3 trp359ala Kd = 1.06 E-4 M; Talin F3 leu325arg Kd= 2.12 E-7 M