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 for Signal-Sequence Recognition by the Translocase Motor SecA as Determined by NMR. Structural Basis for Signal-Sequence Recognition by the Translocase Motor SecA as Determined by NMR. Cell (0092-8674) 2007 imex curation Gelis I., Bonvin AM., Keramisanou D., Koukaki M., Gouridis G., Karamanou S., Economou A., Kalodimos CG. Spectra were recorded at 25C at which temperature SecA is dimeric babis@rutgers.edu 01-April-2008: Contacted by JYOTI. Only protein-protein interactions 5 in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Gelis I., Bonvin AM., Keramisanou D., Koukaki M., Gouridis G., Karamanou S., Economou A., Kalodimos CG. Spectra were recorded at 25C at which temperature SecA is dimeric babis@rutgers.edu Accepted 2008-FEB-28 by CDEROW. 01-April-2008: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 5 imex curation Structural Basis for Signal-Sequence Recognition by the Translocase Motor SecA as Determined by NMR. Structural Basis for Signal-Sequence Recognition by the Translocase Motor SecA as Determined by NMR. Cell (0092-8674) 2007 imex curation Gelis I., Bonvin AM., Keramisanou D., Koukaki M., Gouridis G., Karamanou S., Economou A., Kalodimos CG. Spectra were recorded at 25C at which temperature SecA is dimeric babis@rutgers.edu 01-April-2008: Contacted by JYOTI. Only protein-protein interactions 5 in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Gelis I., Bonvin AM., Keramisanou D., Koukaki M., Gouridis G., Karamanou S., Economou A., Kalodimos CG. Spectra were recorded at 25C at which temperature SecA is dimeric babis@rutgers.edu Accepted 2008-FEB-28 by CDEROW. 01-April-2008: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 5 imex curation Structural Basis for Signal-Sequence Recognition by the Translocase Motor SecA as Determined by NMR. Structural Basis for Signal-Sequence Recognition by the Translocase Motor SecA as Determined by NMR. Cell (0092-8674) 2007 imex curation Gelis I., Bonvin AM., Keramisanou D., Koukaki M., Gouridis G., Karamanou S., Economou A., Kalodimos CG. Spectra were recorded at 25C at which temperature SecA is dimeric babis@rutgers.edu 01-April-2008: Contacted by JYOTI. Only protein-protein interactions 5 in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Gelis I., Bonvin AM., Keramisanou D., Koukaki M., Gouridis G., Karamanou S., Economou A., Kalodimos CG. Spectra were recorded at 25C at which temperature SecA is dimeric babis@rutgers.edu Accepted 2008-FEB-28 by CDEROW. 01-April-2008: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 5 imex curation ppb_ecoli Alkaline phosphatase phoA b0383 JW0374 protein protein ecoli Escherichia coli (strain K12) MKQSTIALALLPLLFTPVTKARTPEMPVLENRAAQGDITAPGGARRLTGDQTAALRDSLSDKPAKNIILLIGDGMGDSEITAARNYAEGAGGFFKGIDALPLTGQYTHYALNKKTGKPDYVTDSAASATAWSTGVKTYNGALGVDIHEKDHPTILEMAKAAGLATGNVSTAELQDATPAALVAHVTSRKCYGPSATSEKCPGNALEKGGKGSITEQLLNARADVTLGGGAKTFAETATAGEWQGKTLREQAQARGYQLVSDAASLNSVTEANQQKPLLGLFADGNMPVRWLGPKATYHGNIDKPAVTCTPNPQRNDSVPTLAQMTDKAIELLSKNEKGFFLQVEGASIDKQDHAANPCGQIGETVDLDEAVQRALEFAKKEGNTLVIVTADHAHASQIVAPDTKAPGLTQALNTKDGAVMVMSYGNSEEDSQEHTGSQLRIAAYGPHAANVVGLTDQTDLFYTMKAALGLK DIP protein P00634 original sequence version: 122 DIP protein P00634 original sequence version: 123 DIP protein P00634 original sequence version: null 8A8DE1F29D9D9253 seca_ecoli Protein translocase subunit SecA secA azi pea prlD b0098 JW0096 protein protein ecoli Escherichia coli (strain K12) MLIKLLTKVFGSRNDRTLRRMRKVVNIINAMEPEMEKLSDEELKGKTAEFRARLEKGEVLENLIPEAFAVVREASKRVFGMRHFDVQLLGGMVLNERCIAEMRTGEGKTLTATLPAYLNALTGKGVHVVTVNDYLAQRDAENNRPLFEFLGLTVGINLPGMPAPAKREAYAADITYGTNNEYGFDYLRDNMAFSPEERVQRKLHYALVDEVDSILIDEARTPLIISGPAEDSSEMYKRVNKIIPHLIRQEKEDSETFQGEGHFSVDEKSRQVNLTERGLVLIEELLVKEGIMDEGESLYSPANIMLMHHVTAALRAHALFTRDVDYIVKDGEVIIVDEHTGRTMQGRRWSDGLHQAVEAKEGVQIQNENQTLASITFQNYFRLYEKLAGMTGTADTEAFEFSSIYKLDTVVVPTNRPMIRKDLPDLVYMTEAEKIQAIIEDIKERTAKGQPVLVGTISIEKSELVSNELTKAGIKHNVLNAKFHANEAAIVAQAGYPAAVTIATNMAGRGTDIVLGGSWQAEVAALENPTAEQIEKIKADWQVRHDAVLEAGGLHIIGTERHESRRIDNQLRGRSGRQGDAGSSRFYLSMEDALMRIFASDRVSGMMRKLGMKPGEAIEHPWVTKAIANAQRKVESRNFDIRKQLLEYDDVANDQRRAIYSQRNELLDVSDVSETINSIREDVFKATIDAYIPPQSLEEMWDIPGLQERLKNDFDLDLPIAEWLDKEPELHEETLRERILAQSIEVYQRKEEVVGAEMMRHFEKGVMLQTLDSLWKEHLAAMDYLRQGIHLRGYAQKDPKQEYKRESFSMFAAMLESLKYEVISTLSKVQVRMPEEVEELEQQRRMEAERLAQMQQLSHQDDDSAAAAALAAQTGERKVGRNDPCPCGSGKKYKQCHGRLQ DIP protein P10408 original sequence version: 150 DIP protein P10408 original sequence version: 130 DIP protein P10408 original sequence version: 105 DIP protein P10408 original sequence version: 107 DIP protein P10408 original sequence version: 125 DIP protein P10408 original sequence version: 140 DIP protein P10408 original sequence version: null 683D66DD4305DBC2 lamb_ecoli Maltoporin lamB malB Maltose-inducible porin b4036 JW3996 Maltose outer membrane channel Phage lambda receptor protein protein protein ecoli Escherichia coli (strain K12) MMITLRKLPLAVAVAAGVMSAQAMAVDFHGYARSGIGWTGSGGEQQCFQTTGAQSKYRLGNECETYAELKLGQEVWKEGDKSFYFDTNVAYSVAQQNDWEATDPAFREANVQGKNLIEWLPGSTIWAGKRFYQRHDVHMIDFYYWDISGPGAGLENIDVGFGKLSLAATRSSEAGGSSSFASNNIYDYTNETANDVFDVRLAQMEINPGGTLELGVDYGRANLRDNYRLVDGASKDGWLFTAEHTQSVLKGFNKFVVQYATDSMTSQGKGLSQGSGVAFDNEKFAYNINNNGHMLRILDHGAISMGDNWDMMYVGMYQDINWDNDNGTKWWTVGIRPMYKWTPIMSTVMEIGYDNVESQRTGDKNNQYKITLAQQWQAGDSIWSRPAIRVFATYAKWDEKWGYDYTGNADNNANFGKAVPADFNGGSFGRGDSDEWTFGAQMEIWW DIP protein P02943 original sequence version: 138 B2D4E4F120DA494E seca-lamb-1 1 5 unspecified role unspecified role neutral component neutral component region 13c label 13c label C13 13C C13 13C undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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) 6 unspecified role unspecified role neutral component neutral component signal peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction The lowest-energy structure of SecA bound to the KRR-LamB signal peptide seca-lamb-2 3 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 P10408:p.[Ile304Ala;Leu306Ala] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain I A certain certain sequence position certain certain certain sequence position certain L A Wed May 02 15:04:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ile304ala,leu306ala' Tue Jun 30 20:05:28 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Ile304Ala;Leu306Ala]' ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role neutral component neutral component signal peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P02943:p.Lys7_Leu8insArgArg mutation increasing mutation increasing interaction mutation increasing insertion analysis insertion analysis certain certain sequence position certain certain certain sequence position certain KL KRRL Wed May 02 18:00:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys7lys_arg_arg' Tue Jun 30 16:11:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys7_Leu8insArgArg' chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Binding isotherms of the calorimetric titration of the KRR-LamB signal peptide to SecA Fig. 4C, 4D, 5C, 5D, S3 Kd of full length SecA binding to insertion mutation at lys-7 of LamB =3E-6M Kd of full length SecA binding to wild type peptide =1E-4M Kd of 1-834 SecA binding to wild type peptide ~ 3E-6M Kd of 1-834 SecA binding to insertion mutation at lys-7 of LamB = 3E-7M iso304ala_leu306ala mutant suggests hydrophobic interactions contribute to signal-peptide recognition by SecA, altering the buffer conditions showed electrostatic interactions to also be important. In the presence of SecB, LamB signal peptide binds to SecA with a much higher affinity (7-fold increase). The D-enantiomer of LamB binds with a lower affinity that the L-enantiomer. seca-phoa-1 3 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 ecobb Escherichia coli (strain B / BL21) 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 chemical synthesis Chemical synthesis (Chemical synthesis) 25 23 direct interaction direct interaction Binding Inhibition of the Alkaline Phosphatase Signal Peptide to SecA Supp. Fig. 11 seca-lamb-4 2 5 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) 6 unspecified role unspecified role neutral component neutral component P02943:p.Lys7_Leu8insArgArg mutation mutation insertion analysis insertion analysis certain certain sequence position certain certain certain sequence position certain KL KRRL Wed May 02 19:47:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys7lys_arg_arg' Tue Jun 30 19:26:16 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys7_Leu8insArgArg' signal peptide sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction Supp Fig. 13 seca-1 2 5 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction Supp Fig. 13