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/ Divergence in the plasminogen-binding group a streptococcal M protein family: functional conservation of binding site and potential role for immune selection of variants. Divergence in the plasminogen-binding group a streptococcal M protein family: functional conservation of binding site and potential role for immune selection of variants. J. Biol. Chem. (0021-9258) 2006 imex curation Sanderson-Smith M., Batzloff M., Sriprakash KS., Dowton M., Ranson M., Walker MJ. BioCreative - Critical Assessment of Information Extraction systems in Biology Assays were carried out in the presence or absence of 50 fold molar excess of unlabelled Glu-plasminogen (UniProt accession number Q5TEH4) or unlabelled fluid phase PAM(NS13) (UniProt accession number Q6V4L5). mwalker@uow.edu.au 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay predetermined predetermined participant predetermined Sanderson-Smith M., Batzloff M., Sriprakash KS., Dowton M., Ranson M., Walker MJ. BioCreative - Critical Assessment of Information Extraction systems in Biology Assays were carried out in the presence or absence of 50 fold molar excess of unlabelled Glu-plasminogen (UniProt accession number Q5TEH4) or unlabelled fluid phase PAM(NS13) (UniProt accession number Q6V4L5). mwalker@uow.edu.au Accepted 2006-AUG-25 by ORCHARD. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Divergence in the plasminogen-binding group a streptococcal M protein family: functional conservation of binding site and potential role for immune selection of variants. Divergence in the plasminogen-binding group a streptococcal M protein family: functional conservation of binding site and potential role for immune selection of variants. J. Biol. Chem. (0021-9258) 2006 imex curation Sanderson-Smith M., Batzloff M., Sriprakash KS., Dowton M., Ranson M., Walker MJ. BioCreative - Critical Assessment of Information Extraction systems in Biology Assays were carried out in the presence or absence of 50 fold molar excess of unlabelled Glu-plasminogen (UniProt accession number Q5TEH4) or unlabelled fluid phase PAM(NS13) (UniProt accession number Q6V4L5). mwalker@uow.edu.au 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro competition binding competition binding predetermined predetermined participant predetermined Sanderson-Smith M., Batzloff M., Sriprakash KS., Dowton M., Ranson M., Walker MJ. BioCreative - Critical Assessment of Information Extraction systems in Biology Assays were carried out in the presence or absence of 50 fold molar excess of unlabelled Glu-plasminogen (UniProt accession number Q5TEH4) or unlabelled fluid phase PAM(NS13) (UniProt accession number Q6V4L5). mwalker@uow.edu.au Accepted 2006-AUG-25 by ORCHARD. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Human C4b-binding protein, structural basis for interaction with streptococcal M protein, a major bacterial virulence factor. Human C4b-binding protein, structural basis for interaction with streptococcal M protein, a major bacterial virulence factor. J. Biol. Chem. (0021-9258) 2006 imex curation Jenkins HT., Mark L., Ball G., Persson J., Lindahl G., Uhrin D., Blom AM., Barlow PN. BioCreative - Critical Assessment of Information Extraction systems in Biology Paul.Barlow@ed.ac.uk 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro saturation binding saturation binding immunostaining immunostaining Jenkins HT., Mark L., Ball G., Persson J., Lindahl G., Uhrin D., Blom AM., Barlow PN. BioCreative - Critical Assessment of Information Extraction systems in Biology Paul.Barlow@ed.ac.uk Accepted 2006-SEP-7 by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Human C4b-binding protein, structural basis for interaction with streptococcal M protein, a major bacterial virulence factor. Human C4b-binding protein, structural basis for interaction with streptococcal M protein, a major bacterial virulence factor. J. Biol. Chem. (0021-9258) 2006 imex curation Jenkins HT., Mark L., Ball G., Persson J., Lindahl G., Uhrin D., Blom AM., Barlow PN. BioCreative - Critical Assessment of Information Extraction systems in Biology Paul.Barlow@ed.ac.uk 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Jenkins HT., Mark L., Ball G., Persson J., Lindahl G., Uhrin D., Blom AM., Barlow PN. BioCreative - Critical Assessment of Information Extraction systems in Biology Paul.Barlow@ed.ac.uk Accepted 2006-SEP-7 by JYOTI. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Human C4b-binding protein, structural basis for interaction with streptococcal M protein, a major bacterial virulence factor. Human C4b-binding protein, structural basis for interaction with streptococcal M protein, a major bacterial virulence factor. J. Biol. Chem. (0021-9258) 2006 imex curation Jenkins HT., Mark L., Ball G., Persson J., Lindahl G., Uhrin D., Blom AM., Barlow PN. BioCreative - Critical Assessment of Information Extraction systems in Biology Paul.Barlow@ed.ac.uk 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Jenkins HT., Mark L., Ball G., Persson J., Lindahl G., Uhrin D., Blom AM., Barlow PN. BioCreative - Critical Assessment of Information Extraction systems in Biology NMR spectra were acquired (37 �C) on Bruker AVANCE 600 and 800 MHz spectrometers. Paul.Barlow@ed.ac.uk Accepted 2006-SEP-7 by JYOTI. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation q6v4l4_strpy protein protein strpy Streptococcus pyogenes RKLKTGTASVAVALTVVGAGLASQTEVKADQPVDHHRYTEANNAVLQGRTVSARALLHEINKNGQLRSENEELKADLQKKEQELKNLNDDVKKLNDEVALERLKNERHVHDEEVELERLKNERHDHDKKEAERKALEDKLADKQEHLDGALRYINEKEAERKEKEAEQKKLKEEKQISDASRQGLRRDLDASREAKKQVEKDLANLTAELDKAKEEKQISDASRQGLRRDLDASREAKKQVEKDLANLTAELDKVKEEKQISDASRQGLRRDLDASREAKKQVEKALEEANSKLAALEKLNKELEESKKLTEKEKAELQAKLEAEAKALKEKLAKQAEELAKLRAEKASDSQTPDAKPGNKAVPGKGQAPQAGTKPNQNKAPMKETKRQLPSTGETT 2EC2C724ABBF8D7C arp4_strpy IgA receptor arp4 protein protein strpy Streptococcus pyogenes MARKDTNKQYSLRKLKTGTASVAVAVAVLGAGFANQTEVKAAEIKKPQADSAWNWPKEYNALLKENEELKVEREKYLSYADDKEKDPQYRALMGENQDLRKREGQYQDKIEELEKERKEKQERQEQLERQYQIEADKHYQEQQKKHQQEQQQLEAEKQKLAKDKQISDASRQGLSRDLEASRAAKKELEAEHQKLKEEKQISDASRQGLSRDLEASREAKKKVEADLAALTAEHQKLKEDKQISDASRQGLSRDLEASREAKKKVEADLAEANSKLQALEKLNKELEEGKKLSEKEKAELQARLEAEAKALKEQLAKQAEELAKLKGNQTPNAKVAPQANRSRSAMTQQKRTLPSTGETANPFFTAAAATVMVSAGMLALKRKEEN C5FBDCDBB97BA0B8 plmn_human Plasminogen PLG protein protein human Homo sapiens Human MEHKEVVLLLLLFLKSGQGEPLDDYVNTQGASLFSVTKKQLGAGSIEECAAKCEEDEEFTCRAFQYHSKEQQCVIMAENRKSSIIIRMRDVVLFEKKVYLSECKTGNGKNYRGTMSKTKNGITCQKWSSTSPHRPRFSPATHPSEGLEENYCRNPDNDPQGPWCYTTDPEKRYDYCDILECEEECMHCSGENYDGKISKTMSGLECQAWDSQSPHAHGYIPSKFPNKNLKKNYCRNPDRELRPWCFTTDPNKRWELCDIPRCTTPPPSSGPTYQCLKGTGENYRGNVAVTVSGHTCQHWSAQTPHTHNRTPENFPCKNLDENYCRNPDGKRAPWCHTTNSQVRWEYCKIPSCDSSPVSTEQLAPTAPPELTPVVQDCYHGDGQSYRGTSSTTTTGKKCQSWSSMTPHRHQKTPENYPNAGLTMNYCRNPDADKGPWCFTTDPSVRWEYCNLKKCSGTEASVVAPPPVVLLPDVETPSEEDCMFGNGKGYRGKRATTVTGTPCQDWAAQEPHRHSIFTPETNPRAGLEKNYCRNPDGDVGGPWCYTTNPRKLYDYCDVPQCAAPSFDCGKPQVEPKKCPGRVVGGCVAHPHSWPWQVSLRTRFGMHFCGGTLISPEWVLTAAHCLEKSPRPSSYKVILGAHQEVNLEPHVQEIEVSRLFLEPTRKDIALLKLSSPAVITDKVIPACLPSPNYVVADRTECFITGWGETQGTFGAGLLKEAQLPVIENKVCNRYEFLNGRVQSTELCAGHLAGGTDSCQGDSGGPLVCFEKDKYILQGVTSWGLGCARPNKPGVYVRVSRFVTWIEGVMRNN 8B31CB877CCB3AB6 q6v4l5_strpy protein protein strpy Streptococcus pyogenes RKLKTGTASVAVALTVVGAGLASQTEVKANRADDARNEVLRGNLVRAELWYRQIQENDQLKLENKGLKTDLREKEEELQGLKDDVEKLTADAELQRLKNERHEEAELERLKSERHDHDKKEAERKALEDKLADKQEHLNGALRYINEKEAERKEKEAEQKKLKEEKQISDASRQGLRRDLDASREAKKQVEKDLANLTAELDKVKEEKQISDASRQGLRRDLDASREAKKQVEKGLANLTAELDKVKEEKQISDASRQGLRRDLDASREAKKQVEKALEEANSKLAALEKLNKELEESKKLTEKEKAELQAKLEAEAKALKEQLAKQAEELAKLRAEKASDSQTPDAKPGNKAVPGKGQAPQAGTKPNQNKAPMKETKRQLPSTSETT 2B3722727C5002DF c4bpa_human C4b-binding protein alpha chain Proline-rich protein C4BPA C4BP protein protein human Homo sapiens Human MHPPKTPSGALHRKRKMAAWPFSRLWKVSDPILFQMTLIAALLPAVLGNCGPPPTLSFAAPMDITLTETRFKTGTTLKYTCLPGYVRSHSTQTLTCNSDGEWVYNTFCIYKRCRHPGELRNGQVEIKTDLSFGSQIEFSCSEGFFLIGSTTSRCEVQDRGVGWSHPLPQCEIVKCKPPPDIRNGRHSGEENFYAYGFSVTYSCDPRFSLLGHASISCTVENETIGVWRPSPPTCEKITCRKPDVSHGEMVSGFGPIYNYKDTIVFKCQKGFVLRGSSVIHCDADSKWNPSPPACEPNSCINLPDIPHASWETYPRPTKEDVYVVGTVLRYRCHPGYKPTTDEPTTVICQKNLRWTPYQGCEALCCPEPKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETSRFSAICQGDGTWSPRTPSCGDICNFPPKIAHGHYKQSSSYSFFKEEIIYECDKGYILVGQAKLSCSYSHWSAPAPQCKALCRKPELVNGRLSVDKDQYVEPENVTIQCDSGYGVVGPQSITCSGNRTWYPEVPKCEWETPEGCEQVLTGKRLMQCLPNPEDVKMALEVYKLSLEIEQLELQRDSARQSTLDKEL 67E03F2EA85A16DD q6v4l1_strpy protein protein strpy Streptococcus pyogenes RKLKTGTASVAVALTVVGAGLASQTEVKAENHPGYTAAQNGVLSELPGQAQAFSRAFLHEREKNGELRLENEGLKTDLKKKEQELESLKGDVEKLKDEVALERLKNERHVHDEEAELERLKNERYEHDKKEAERKALEDKLADKQEHLNGALRYINEKEAERKEKEAEQKKLKEEKQISDASRQGLRRDLDASREAKKQVEKDLANLTAELDKVKEEKQVSDASRQGLRRDLDASREAKKKVEADLAALTAEHQKLKEEKQISDASRQGLSRDLEASREAKKKVEADLAEANSKLQALEKLNKELEEGKKLSEKEKAELQARLEAEAKALKEQLAKQAEELAKLRAGKASDSQTPDAKPGNKAVPGKGQAPQAGTKPNQNKAPMKETKRQLPSTGETT 00C3CC24C19BD311 q6v4k9_strpy protein protein strpy Streptococcus pyogenes RKLKTGTASVAVALTVVGAGLASQTEVKADQPVDHHRYTEANNAVLQGRTVSARALLHEINKNGQLRSENEELKADLQKKEQELKNLNDDVKKLNDEVALERLKNERHVHDEEVELERLKNERHDHDKKEAERKALEDKLADKQEHLDGALRYINEKEAERKEKEAEQKKLKEEKQISDASRQGLRRDLDASREAKKQVEKALEEANSKLAALEKLNKELEESKKLTEKEKAELQAKLEAEAKALKEKLAKQAEELAKLRAEKASDSQTPDAKPGNKAVPGKGQAPQAGTKPNQNKAPMKETKRQLPSTG 253174D4F7790093 q6v4l9_strpy protein protein strpy Streptococcus pyogenes RKLKTGTASVAVALTVLGAGFANQTEVKAADHPSYTAAKDEVLSKFSVPGHVWAHEREKNDKLSSENEGLKAGLQEKEQELKNLKADVEKLKDAAELERLKNERHDHDELERLKNERHDHDKREAERKALEDKLADKQEHLDGALRYINEKEAERKEKEAEQKKLKEEKQISDASRQGLRRDLDASREAKKQVEKDLANLTAELDKVKEEKQISDASRQGLRRDLDASREAKKQVEKDLADLTAELDKVKEEKQISDASRQGLRRDLDASREAKKQVEKALEEANSKLAALEKLNKELEESKKLTEKEKAELQAKLEAEAKALKEQLAKQAEELAKLRAEKASDSQTPDAKPGNKAVPGKGQAPQAGTKPNQNKAPMKETKRQLPSTGETT 022DB5B0DA016B88 apr4-c4bp 3 7 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 10 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 1A To define the C4BP region involved in M protein binding, deletion mutants of the -chain were tested for binding to theMprotein, apr4-c4bp-1 3 10 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 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 1A To define the C4BP region involved in M protein binding, deletion mutants of the -chain were tested for binding to theMprotein, apr4-c4bp-2 3 10 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 1B In further investigations, the linking sequences between CCPs 1, 2, 3, and 4 of the -chain were engineered by insertions of Ala residues (Fig. 1B). Addition of residues between CCP2 and CCP3, or between CCP3 and CCP4, had no effect on binding, whereas insertions between CCP1 and CCP2 caused a loss of M4 binding. Insertion of two alanine residues between Lysine 188 and Isoleucine 189 resulted in abolished interaction between the two proteins. apr4-c4bp-3 4 10 unspecified role unspecified role neutral component neutral component c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 1C A recombinant protein consisting of residues 1�124 of C4BP -chain (see Fig. 1C, C4BP12), His-tagged at the C terminus, was expressed in E. coli and refolded. ITC was used (Fig. 1D) to measure the binding affinity between C4BP12 and a dimerised peptide corresponding to the hypervariable region of M4 (residues 1�45 of M4 protein with a C-terminal Cys added. A dimer of the APR4 protein was created as previous studies had shown that dimerization strongly enhanced C4BP binding. The APR4 protein used in this stidy also had a C-terminal Cys added. apr4-c4bp-4 5 10 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 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role neutral component neutral component n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 4 and Figure 5 Significant changes in the 15N-1H HSQC spectrum of C4BP12 occurred upon addition of M4-N The APR4 protein was present as a dimer. plg-q6v4l5 2 8 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined glu N-terminal Glu residue experimental feature experimental feature n-term range n-terminal range n-term range n-term range n-terminal range n-term range 9 unspecified role unspecified role bait bait c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 3 Figure 4 Table 2 The circulating form of plasminogen was isolated from Human plasma which has a Glutamic acid residue attached to the N-termini of the plasminogen protein. Effective concentration of competitor (unlabelled Q6V4L5) required to inhibit binding by 50% (EC50) was 22.06 micro Molar. Saturation binding analysis of biotinylated Glu-plasminogen to immobilized recombinant PAM variant proteins. plg-q6v4l5-1 1 8 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined glu N-terminal Glu residue experimental feature experimental feature n-term range n-terminal range n-term range n-term range n-terminal range n-term range 9 unspecified role unspecified role bait bait c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 2C The circulating form of plasminogen was isolated from Human plasma which has a Glutamic acid residue attached to the N-termini of the plasminogen protein. ligand blot analysis employing biotinylated Glu-plasminogen of purified variant PAM proteins. Lane 1, NS696 M1 protein; lane 2, PAMNS265; lane 3, PAMNS455; lane 4, PAMNS1133; lane 5, PAMNS53; lane 6, PAMNS13 plg-q6v4l9 2 8 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined glu N-terminal Glu residue experimental feature experimental feature n-term range n-terminal range n-term range n-term range n-terminal range n-term range 13 unspecified role unspecified role bait bait c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 3 Figure 4 Table 2 The circulating form of plasminogen was isolated from Human plasma which has a Glutamic acid residue attached to the N-termini of the plasminogen protein. Effective concentration of competitor (unlabelled Q6V4L9) required to inhibit binding by 50% (EC50) was 0.68 micro Molar. Saturation binding analysis of biotinylated Glu-plasminogen to immobilized recombinant PAM variant proteins. plg-q6v4l9-1 1 13 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined glu N-terminal Glu residue experimental feature experimental feature n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction Figure 2C The circulating form of plasminogen was isolated from Human plasma which has a Glutamic acid residue attached to the N-termini of the plasminogen protein. ligand blot analysis employing biotinylated Glu-plasminogen of purified variant PAM proteins. Lane 1, NS696 M1 protein; lane 2, PAMNS265; lane 3, PAMNS455; lane 4, PAMNS1133; lane 5, PAMNS53; lane 6, PAMNS13 plg-q6v4l1 1 11 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined glu N-terminal Glu residue experimental feature experimental feature n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction Figure 2C The circulating form of plasminogen was isolated from Human plasma which has a Glutamic acid residue attached to the N-termini of the plasminogen protein. ligand blot analysis employing biotinylated Glu-plasminogen of purified variant PAM proteins. Lane 1, NS696 M1 protein; lane 2, PAMNS265; lane 3, PAMNS455; lane 4, PAMNS1133; lane 5, PAMNS53; lane 6, PAMNS13 plg-q6v4l4 2 6 unspecified role unspecified role bait bait c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined glu N-terminal Glu residue experimental feature experimental feature n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction Figure 3 Figure 4 Table 2 The circulating form of plasminogen was isolated from Human plasma which has a Glutamic acid residue attached to the N-termini of the plasminogen protein. Effective concentration of competitor (unlabelled Q6V4L4) required to inhibit binding by 50% (EC50) was 1.96 micro Molar. Saturation binding analysis of biotinylated Glu-plasminogen to immobilized recombinant PAM variant proteins. plg-q6v4l4-1 1 6 unspecified role unspecified role bait bait n-terminal gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined glu N-terminal Glu residue experimental feature experimental feature n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction Figure 2C The circulating form of plasminogen was isolated from Human plasma which has a Glutamic acid residue attached to the N-termini of the plasminogen protein. ligand blot analysis employing biotinylated Glu-plasminogen of purified variant PAM proteins. Lane 1, NS696 M1 protein; lane 2, PAMNS265; lane 3, PAMNS455; lane 4, PAMNS1133; lane 5, PAMNS53; lane 6, PAMNS13 plg-q6v4l1-1 2 11 unspecified role unspecified role bait bait c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined glu N-terminal Glu residue experimental feature experimental feature n-term range n-terminal range n-term range n-term range n-terminal range n-term range direct interaction direct interaction Figure 3 Figure 4 Table 2 The circulating form of plasminogen was isolated from Human plasma which has a Glutamic acid residue attached to the N-termini of the plasminogen protein. Effective concentration of competitor (unlabelled Q6V4L1) required to inhibit binding by 50% (EC50) was 0.81 micro Molar. Saturation binding analysis of biotinylated Glu-plasminogen to immobilized recombinant PAM variant proteins. plg-q6v4k9 2 8 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined glu N-terminal Glu residue experimental feature experimental feature n-term range n-terminal range n-term range n-term range n-terminal range n-term range 12 unspecified role unspecified role bait bait c terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) direct interaction direct interaction Figure 3 Figure 4 Table 2 The circulating form of plasminogen was isolated from Human plasma which has a Glutamic acid residue attached to the N-termini of the plasminogen protein. Effective concentration of competitor (unlabelled Q6V4K9) required to inhibit binding by 50% (EC50) was 5.77 micro Molar. Saturation binding analysis of biotinylated Glu-plasminogen to immobilized recombinant PAM variant proteins. mpidb The Microbial Protein Interaction Database In vivo interactions between toxin-antitoxin proteins epsilon and zeta of streptococcal plasmid pSM19035 in Saccharomyces cerevisiae. In vivo interactions between toxin-antitoxin proteins epsilon and zeta of streptococcal plasmid pSM19035 in Saccharomyces cerevisiae. J. Bacteriol. (0021-9193) 2009 imex curation Zielenkiewicz U., Kowalewska M., Kaczor C., Ceglowski P. ulazet@ibb.waw.pl Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Zielenkiewicz U., Kowalewska M., Kaczor C., Ceglowski P. ulazet@ibb.waw.pl Accepted 2013-FEB-08 by ORCHARD J. Bacteriol. (0021-9193) 2009 Only protein-protein interactions imex curation ztox_strpy Toxin zeta UDP-N-acetylglucosamine kinase protein protein strpy Streptococcus pyogenes MANIVNFTDKQFENRLNDNLEELIQGKKAVESPTAFLLGGQPGSGKTSLRSAIFEETQGNVIVIDNDTFKQQHPNFDELVKLYEKDVVKHVTPYSNRMTEAIISRLSDQGYNLVIEGTGRTTDVPIQTATMLQAKGYETKMYVMAVPKINSYLGTIERYETMYADDPMTARATPKQAHDIVVKNLPTNLETLHKTGLFSDIRLYNREGVKLYSSLETPSISPKETLEKELNRKVSGKEIQPTLERIEQKMVLNKHQETPEFKAIQQKLESLQPPTPPIPKTPKLPGI DIP protein Q54944 original sequence version: 61 B0C6282EE5B2210F eatx_strpy Antitoxin epsilon protein protein strpy Streptococcus pyogenes MAVTYEKTFEIEIINELSASVYNRVLNYVLNHELNKNDSQLLEVNLLNQLKLAKRVNLFDYSLEELQAVHEYWRSMNRYSKQVLNKEKVA DIP protein Q57231 original sequence version: 53 31628C7F2FDE4B50 ztox-eatx 1 2 unspecified role unspecified role bait bait 3 unspecified role unspecified role prey prey region Q57231-1-12 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association table 3, 4 ztox-eatx-1 1 3 unspecified role unspecified role prey prey region Q57231-1-66 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region Q57231-1-29 required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 2 unspecified role unspecified role bait bait physical association physical association table 3, 4 eatx-ztox 1 3 unspecified role unspecified role bait bait 2 unspecified role unspecified role prey prey physical association physical association table 3 MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it A novel adaptation of aldolase regulates virulence in Streptococcus pyogenes. A novel adaptation of aldolase regulates virulence in Streptococcus pyogenes. EMBO J. (0261-4189) 2006 imex curation Loughman JA., Caparon MG. 2009-01-08 Only protein-protein interactions strp1 Streptococcus pyogenes serotype M1 anti tag coip anti tag coimmunoprecipitation anti tag coip nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Loughman JA., Caparon MG. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation o85731_strpy ropB protein protein strpy Streptococcus pyogenes MEIGETVEFIRHSKNISIKQVCGDYLTRQTYYRFIKNNLDISSKKLLYILDNLNVNVDEFLFISNNFKQYKEFIDMDTAKHYFECRNIEGLNHILDSYKDSKSTKEKNLFALVKVLLATLTEEDCLTERTYLSNYLINIETWSHYETVLFNNCMFILESCFIEMVFSKVIVNLDKYNTLRYYGNESIRMFVNMLILFIQRQEYDKASEILAKIEDYQLNDDCLYERCCVSFFDGIIGLINGKEGAEQKCVQILEIFQLLNCKTIHHMFQTYLEAIKHKLS mint 85DA9ADE019092CD lacd1_strp1 Tagatose 1,6-diphosphate aldolase 1 SPy_1704 lacD1 lacD.1 M5005_Spy1395 Tagatose-bisphosphate aldolase 1 D-tagatose-1,6-bisphosphate aldolase 1 protein protein strp1 Streptococcus pyogenes serotype M1 MTITANKRHYLEKVSHQGIISALAFDQRGALKQMMAAHQEGEATVTQIETLKVLVSEELTPYASSILLDPEYGLLATKVRANQTGLLLAYEKTGYDATTTSRLPDCLVEWSVKRLKAAGADAIKFLLYYDVDGDEQINLQKQAYIERIGSECTAEDIPFFLELLSYDERISDNNSAAYAKLKPHKVNGAMSVFSDKRFGVDVLKVEVPVNMAYVEGFTEGEVHYSQAEAIKAFQDQEAASHLPYIYLSAGVSAKLFQETLYFAAAAGAQFSGVLCGRATWAGSVPVYITKGEDEARKWLCTEGFQNIDELNRVLEETASPWTEKI mint 7B3A1B69AF3C471E ropb-lacd1 1 3 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tagged 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 Stoichiometry: 1.0 2 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell ha tagged ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F3B EMBO-Journal mint DIP Database of Interacting Proteins 611 Young Drive East; Los Angeles CA 90095; USA http://dip.doe-mbi.ucla.edu/ dip@mbi.ucla.edu Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Nature 2017 imex curation Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro mst microscale thermophoresis mst predetermined predetermined participant predetermined Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn Accepted 2019-JUL-04 AT 09:18 BST AT 09:18 BST by ORCHARD Nature 2017 imex curation Hi Sandra, I have fixed everything you mentioned, except for the Kds as discussed previously with Lukasz. Thank you! Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Nature 2017 imex curation Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro pull down pull down experimental particp experimental participant identification experimental particp Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn Accepted 2019-JUL-04 AT 09:18 BST AT 09:18 BST by ORCHARD Nature 2017 imex curation Hi Sandra, I have fixed everything you mentioned, except for the Kds as discussed previously with Lukasz. Thank you! Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. Structure (London, England : 1993) 2011 imex curation Smith CL., Ghosh J., Elam JS., Pinkner JS., Hultgren SJ., Caparon MG., Ellenberger T. Only protein-protein interactions ecolx Escherichia coli x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Smith CL., Ghosh J., Elam JS., Pinkner JS., Hultgren SJ., Caparon MG., Ellenberger T. DIP original experiment: DIP-218759X Structure (London, England : 1993) 2011 IMEx Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Nature 2017 imex curation Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction experimental particp experimental participant identification experimental particp Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn Accepted 2019-JUL-04 AT 09:18 BST AT 09:18 BST by ORCHARD Nature 2017 imex curation Hi Sandra, I have fixed everything you mentioned, except for the Kds as discussed previously with Lukasz. Thank you! Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Structural basis of CRISPR-SpyCas9 inhibition by an anti-CRISPR protein. Nature 2017 imex curation Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn 25-Jul-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column experimental particp experimental participant identification experimental particp Dong null., Guo M., Wang S., Zhu Y., Wang S., Xiong Z., Yang J., Xu Z., Huang Z. huangzhiwei@hit.edu.cn Accepted 2019-JUL-04 AT 09:18 BST AT 09:18 BST by ORCHARD Nature 2017 imex curation Hi Sandra, I have fixed everything you mentioned, except for the Kds as discussed previously with Lukasz. Thank you! Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. Structure (London, England : 1993) 2011 imex curation Smith CL., Ghosh J., Elam JS., Pinkner JS., Hultgren SJ., Caparon MG., Ellenberger T. Only protein-protein interactions ecolx Escherichia coli molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Smith CL., Ghosh J., Elam JS., Pinkner JS., Hultgren SJ., Caparon MG., Ellenberger T. DIP original experiment: DIP-218758X Structure (London, England : 1993) 2011 IMEx Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. A novel mechanism of programmed cell death in bacteria by toxin-antitoxin systems corrupts peptidoglycan synthesis. A novel mechanism of programmed cell death in bacteria by toxin-antitoxin systems corrupts peptidoglycan synthesis. PLoS biology 2011 imex curation Mutschler H., Gebhardt M., Shoeman RL., Meinhart A. Only protein-protein interactions ecolx Escherichia coli x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Mutschler H., Gebhardt M., Shoeman RL., Meinhart A. DIP original experiment: DIP-216767X PLoS biology 2011 IMEx A novel mechanism of programmed cell death in bacteria by toxin-antitoxin systems corrupts peptidoglycan synthesis. Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. Structure (London, England : 1993) 2011 imex curation Smith CL., Ghosh J., Elam JS., Pinkner JS., Hultgren SJ., Caparon MG., Ellenberger T. Only protein-protein interactions ecolx Escherichia coli ion exchange chrom ion exchange chromatography IEC IEC ion exchange chrom predetermined predetermined participant predetermined Smith CL., Ghosh J., Elam JS., Pinkner JS., Hultgren SJ., Caparon MG., Ellenberger T. DIP original experiment: DIP-218757X Structure (London, England : 1993) 2011 IMEx Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. Pathogenic bacteria attach to human fibronectin through a tandem beta-zipper. Pathogenic bacteria attach to human fibronectin through a tandem beta-zipper. Nature 2003 imex curation Schwarz-Linek U., Werner JM., Pickford AR., Gurusiddappa S., Kim JH., Pilka ES., Briggs JA., Gough TS., Höök M., Campbell ID., Potts JR. jennifer.potts@bioch.ox.ac.uk 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Schwarz-Linek U., Werner JM., Pickford AR., Gurusiddappa S., Kim JH., Pilka ES., Briggs JA., Gough TS., Höök M., Campbell ID., Potts JR. jennifer.potts@bioch.ox.ac.uk Accepted 2018-DEC-13 AT 11:20 GMT AT 11:20 GMT by ORCHARD Nature 2003 imex curation Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. Structure (London, England : 1993) 2011 imex curation Smith CL., Ghosh J., Elam JS., Pinkner JS., Hultgren SJ., Caparon MG., Ellenberger T. Only protein-protein interactions ecolx Escherichia coli pull down pull down predetermined predetermined participant predetermined Smith CL., Ghosh J., Elam JS., Pinkner JS., Hultgren SJ., Caparon MG., Ellenberger T. DIP original experiment: DIP-218756X Structure (London, England : 1993) 2011 IMEx Structural basis of Streptococcus pyogenes immunity to its NAD+ glycohydrolase toxin. a0a459ndx2_lismn acrIIA2 D4271_13320 D4D89_13255 B1N40_08540 B4X79_13415 E0I39_13150 FJU19_12475 G3R95_002503 AF006_07510 APD94_09090 D4C60_14490 D4D22_14105 D5M63_13385 E1V33_15495 protein protein lismn Listeria monocytogenes MTLTRAQKKYAEAMHEFINMVDDFEESTPDFAKEVLHDSDYVVITKNEKYAVALCSLSTDECEYDTNLYLDEKLVDYSTVDVNGVTYYINIVETNDIDDLEIATDEDEMKSGNQEIILKSELK 47F691CB7CD8F4E3 finc_human Fibronectin FN1 FN Cold-insoluble globulin protein protein human Homo sapiens Human MLRGPGPGLLLLAVQCLGTAVPSTGASKSKRQAQQMVQPQSPVAVSQSKPGCYDNGKHYQINQQWERTYLGNALVCTCYGGSRGFNCESKPEAEETCFDKYTGNTYRVGDTYERPKDSMIWDCTCIGAGRGRISCTIANRCHEGGQSYKIGDTWRRPHETGGYMLECVCLGNGKGEWTCKPIAEKCFDHAAGTSYVVGETWEKPYQGWMMVDCTCLGEGSGRITCTSRNRCNDQDTRTSYRIGDTWSKKDNRGNLLQCICTGNGRGEWKCERHTSVQTTSSGSGPFTDVRAAVYQPQPHPQPPPYGHCVTDSGVVYSVGMQWLKTQGNKQMLCTCLGNGVSCQETAVTQTYGGNSNGEPCVLPFTYNGRTFYSCTTEGRQDGHLWCSTTSNYEQDQKYSFCTDHTVLVQTRGGNSNGALCHFPFLYNNHNYTDCTSEGRRDNMKWCGTTQNYDADQKFGFCPMAAHEEICTTNEGVMYRIGDQWDKQHDMGHMMRCTCVGNGRGEWTCIAYSQLRDQCIVDDITYNVNDTFHKRHEEGHMLNCTCFGQGRGRWKCDPVDQCQDSETGTFYQIGDSWEKYVHGVRYQCYCYGRGIGEWHCQPLQTYPSSSGPVEVFITETPSQPNSHPIQWNAPQPSHISKYILRWRPKNSVGRWKEATIPGHLNSYTIKGLKPGVVYEGQLISIQQYGHQEVTRFDFTTTSTSTPVTSNTVTGETTPFSPLVATSESVTEITASSFVVSWVSASDTVSGFRVEYELSEEGDEPQYLDLPSTATSVNIPDLLPGRKYIVNVYQISEDGEQSLILSTSQTTAPDAPPDTTVDQVDDTSIVVRWSRPQAPITGYRIVYSPSVEGSSTELNLPETANSVTLSDLQPGVQYNITIYAVEENQESTPVVIQQETTGTPRSDTVPSPRDLQFVEVTDVKVTIMWTPPESAVTGYRVDVIPVNLPGEHGQRLPISRNTFAEVTGLSPGVTYYFKVFAVSHGRESKPLTAQQTTKLDAPTNLQFVNETDSTVLVRWTPPRAQITGYRLTVGLTRRGQPRQYNVGPSVSKYPLRNLQPASEYTVSLVAIKGNQESPKATGVFTTLQPGSSIPPYNTEVTETTIVITWTPAPRIGFKLGVRPSQGGEAPREVTSDSGSIVVSGLTPGVEYVYTIQVLRDGQERDAPIVNKVVTPLSPPTNLHLEANPDTGVLTVSWERSTTPDITGYRITTTPTNGQQGNSLEEVVHADQSSCTFDNLSPGLEYNVSVYTVKDDKESVPISDTIIPEVPQLTDLSFVDITDSSIGLRWTPLNSSTIIGYRITVVAAGEGIPIFEDFVDSSVGYYTVTGLEPGIDYDISVITLINGGESAPTTLTQQTAVPPPTDLRFTNIGPDTMRVTWAPPPSIDLTNFLVRYSPVKNEEDVAELSISPSDNAVVLTNLLPGTEYVVSVSSVYEQHESTPLRGRQKTGLDSPTGIDFSDITANSFTVHWIAPRATITGYRIRHHPEHFSGRPREDRVPHSRNSITLTNLTPGTEYVVSIVALNGREESPLLIGQQSTVSDVPRDLEVVAATPTSLLISWDAPAVTVRYYRITYGETGGNSPVQEFTVPGSKSTATISGLKPGVDYTITVYAVTGRGDSPASSKPISINYRTEIDKPSQMQVTDVQDNSISVKWLPSSSPVTGYRVTTTPKNGPGPTKTKTAGPDQTEMTIEGLQPTVEYVVSVYAQNPSGESQPLVQTAVTNIDRPKGLAFTDVDVDSIKIAWESPQGQVSRYRVTYSSPEDGIHELFPAPDGEEDTAELQGLRPGSEYTVSVVALHDDMESQPLIGTQSTAIPAPTDLKFTQVTPTSLSAQWTPPNVQLTGYRVRVTPKEKTGPMKEINLAPDSSSVVVSGLMVATKYEVSVYALKDTLTSRPAQGVVTTLENVSPPRRARVTDATETTITISWRTKTETITGFQVDAVPANGQTPIQRTIKPDVRSYTITGLQPGTDYKIYLYTLNDNARSSPVVIDASTAIDAPSNLRFLATTPNSLLVSWQPPRARITGYIIKYEKPGSPPREVVPRPRPGVTEATITGLEPGTEYTIYVIALKNNQKSEPLIGRKKTDELPQLVTLPHPNLHGPEILDVPSTVQKTPFVTHPGYDTGNGIQLPGTSGQQPSVGQQMIFEEHGFRRTTPPTTATPIRHRPRPYPPNVGEEIQIGHIPREDVDYHLYPHGPGLNPNASTGQEALSQTTISWAPFQDTSEYIISCHPVGTDEEPLQFRVPGTSTSATLTGLTRGATYNVIVEALKDQQRHKVREEVVTVGNSVNEGLNQPTDDSCFDPYTVSHYAVGDEWERMSESGFKLLCQCLGFGSGHFRCDSSRWCHDNGVNYKIGEKWDRQGENGQMMSCTCLGNGKGEFKCDPHEATCYDDGKTYHVGEQWQKEYLGAICSCTCFGGQRGWRCDNCRRPGGEPSPEGTTGQSYNQYSQRYHQRTNTNVNCPIECFMPLDVQADREDSRE DIP protein P02751 original sequence version: 119 DIP protein P02751 original sequence version: 131 DIP protein P02751 original sequence version: 154 DIP protein P02751 original sequence version: 156 DIP protein P02751 original sequence version: 157 DIP protein P02751 original sequence version: 155 DIP protein P02751 original sequence version: 194 6C436A7A5FEE6DEB cas9_strp1 CRISPR-associated endonuclease Cas9/Csn1 cas9 csn1 SPy_1046 SpCas9 SpyCas9 protein protein strp1 Streptococcus pyogenes serotype M1 MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD DIP protein Q99ZW2 original sequence version: 77 ztox_strpy Toxin zeta UDP-N-acetylglucosamine kinase protein protein strpy Streptococcus pyogenes MANIVNFTDKQFENRLNDNLEELIQGKKAVESPTAFLLGGQPGSGKTSLRSAIFEETQGNVIVIDNDTFKQQHPNFDELVKLYEKDVVKHVTPYSNRMTEAIISRLSDQGYNLVIEGTGRTTDVPIQTATMLQAKGYETKMYVMAVPKINSYLGTIERYETMYADDPMTARATPKQAHDIVVKNLPTNLETLHKTGLFSDIRLYNREGVKLYSSLETPSISPKETLEKELNRKVSGKEIQPTLERIEQKMVLNKHQETPEFKAIQQKLESLQPPTPPIPKTPKLPGI DIP protein Q54944 original sequence version: 61 B0C6282EE5B2210F a0a247d711_lismn acrIIA4 G3O21_003035 APD94_15065 GON91_02470 UI29_15095 protein protein lismn Listeria monocytogenes MNINDLIREIKNKDYTVKLSGTDSNSITQLIIRVNNDGNEYVISESENESIVEKFISAFKNGWNQEYEDEEEFYNDMQTITLKSELN F44958822FEFF9A5 urs000080e253_strpy_grna URS000080E253 gRNA grna guide rna guide RNA grna strpy Streptococcus pyogenes GGAAAUUAGGUGCGCUUGGCGUUUUAGAGCUAGAAAUAGCAAGUUAAAAUAAGGCUAGUCCGUUAUCAACUUGAAAAAGUGGCACCGAGUCGGUGCUU q5xe42_strp6 M6_Spy0186 protein protein strp6 Streptococcus pyogenes serotype M6 (strain ATCC BAA-946 / MGAS10394) MVYMRNKKVTLAHIVAKTSVAIALAGAMGSSLLANSTTYAVSGKENKKSDVKYETTKVMEANATSSKEDNHVMHTLDGSMSTVWEENSPGGGVGEVLSYKFASPMRIGRILIVNGDTSSKENYYKKNRIAKADVKYYNGNKLVLFQKIELGDTYTKKPHHIEIDKKLDVDRIDIEVTEVHQGQNKDILALSEVTFGNMERDLFEKKFKEIKDKWVTDKQADEFIETADKYADKAVQMSAVASRAEYYRMYVSRKYQYKKEFVEKLKQVYKESGASHVTSKKDLMLAFDDAKRKSTIGRQENGLFVTSFAEDMALLFTDQGKLKSADQIENIKGVDSGKYSDGVYQYEYDSELTKNIDKLGYIRTASGDTPGANSLNIPGCQTWSGKHIENSESELIFPSISVKDLKSKAVLAEIDAKGYFEIIDPTIIAPNGDHKKVTGRFKIKKMQDRMQDRK D9FBF97B708F7828 eatx_strpy Antitoxin epsilon protein protein strpy Streptococcus pyogenes MAVTYEKTFEIEIINELSASVYNRVLNYVLNHELNKNDSQLLEVNLLNQLKLAKRVNLFDYSLEELQAVHEYWRSMNRYSKQVLNKEKVA DIP protein Q57231 original sequence version: 53 31628C7F2FDE4B50 q01924_strpy Sfb protein protein strpy Streptococcus pyogenes MNNKMFLNKEAGFLAHTKRKRRFAVTLVGVFFMLLASAGAIGFGQVAYAADEKTVPHRVSQNPEFPWYGYDFYKGPYTRYHNLQLNLNGSKTYQAYCFNLKRFEPKKEGSYFPNWYKRWDGSEETFVKYADNPRKDNESSRVIDVELEKNILRVLYNGYPNNGNGIMEGLEPLNAILVTQNAVWYYSDNSSIFNTDNFFTTEAKDLNIKPEQLSLMRVALKKLIDPKLSEESLKPVPSTFRLNIFESQDKLYQNLLSAEFVPENPPKPGETPEHGPKTPELDGTPIPEGPQRPNESLEPTLPPVMLDGQEVPEVPSESLEPALPPLMPELDGQEVPEVPSESLEPALPPLMPELDGQEVPEKPSVDLPIEDPRYEFNNKDQSPLAGESGETEYITEVYGNQQNPVDIDKKLPNETGFSGNMVETEDTKEPGVLMGGQSESVEFTKDTQTGMSGQTTPQVETEDTKEPGVLMGGQSESVEFTKDTQTGMSGQTASQVETEDTKEPGVLMGGQSESVEFTKDTQTGMSGQTTPQVETEDTKEPGVLMGGQSESVEFTKDTQTGMSGFSETVTIVEDTRPKLVFHFDNNEPKVEENREKPTKNITPILPATGDIENVLAFLGILILSVLPIFSLLKKQTKQ D55E2A4E5708F3E0 q2vj58_strpy E0F66_06230 ifs GTK54_00720 SPNIH34_01310 protein protein strpy Streptococcus pyogenes MYKVPKGLEHYQKMFQKEVTVNDLKKYLIGSDKEYRITRRDSYMGDISDPEVILEYGVYPAFIKGYTQLKANIEEALLEMSNSGQALDIYQAVQTLNAENMLLNYYESLPFYLNRQSILANITKALKDAHIREAMAHYKLGEFAHYQDTMLDMVERTIETF BEB604A6FB730D7B ifs-spn-1 6 20 unspecified role unspecified role unspecified role unspecified role ecolx Escherichia coli 17 unspecified role unspecified role unspecified role unspecified role C-terminal glycohydrolase domain Region 194-454 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli direct interaction direct interaction spn-ifs-1 10 20 unspecified role unspecified role prey prey ecolx Escherichia coli 17 unspecified role unspecified role bait bait C-terminal glycohydrolase domain Region 194-454 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain His-tag His-tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecolx Escherichia coli physical association physical association ifs-spn-2 3 20 unspecified role unspecified role unspecified role unspecified role ecolx Escherichia coli 17 unspecified role unspecified role unspecified role unspecified role C-terminal glycohydrolase domain Region 194-454 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli direct interaction direct interaction ifs-spn-3 8 20 unspecified role unspecified role unspecified role unspecified role ecolx Escherichia coli 17 unspecified role unspecified role unspecified role unspecified role C-terminal glycohydrolase domain Region 194-454 binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain ecolx Escherichia coli direct interaction direct interaction eatx_strpy-ztox_strpy-1 7 14 unspecified role unspecified role unspecified role unspecified role ecolx Escherichia coli 18 unspecified role unspecified role unspecified role unspecified role ecolx Escherichia coli direct interaction direct interaction fn1-sfb-1 9 12 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain picpa Pichia pastoris (Yeast) Yeast Pichia pastoris 19 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain in vitro In vitro direct interaction direct interaction Fig 3 fn1-sfb-3 9 12 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain picpa Pichia pastoris (Yeast) Yeast Pichia pastoris 19 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain in vitro In vitro direct interaction direct interaction Fig 3 fn1-sfb-4 9 12 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain picpa Pichia pastoris (Yeast) Yeast Pichia pastoris 19 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain in vitro In vitro direct interaction direct interaction Fig 3 fn1-sfb-5 9 12 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain picpa Pichia pastoris (Yeast) Yeast Pichia pastoris 19 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain in vitro In vitro direct interaction direct interaction Fig 3 fn1-sfb-6 9 12 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain picpa Pichia pastoris (Yeast) Yeast Pichia pastoris 19 unspecified role unspecified role neutral component neutral component purified purified binding region binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain in vitro In vitro direct interaction direct interaction Fig 3 a0a247d711_lismn-cas9-1 5 16 weight by bromide molecular weight estimation by bromide staining weight by bromide experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated in vitro In vitro 13 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 15 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) physical association physical association Figure 1 a0a247d711_lismn-cas9-2 4 15 predetermined predetermined participant predetermined structure determination identification by structure determination identification by structure determination structure determination experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 13 predetermined predetermined participant predetermined structure determination identification by structure determination identification by structure determination structure determination experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 16 unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated in vitro In vitro physical association physical association Figure 2, Extended Data Table 1 a0a394vx67_lismn-cas9-1 1 16 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated in vitro In vitro 13 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 11 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) physical association physical association Figure 4d cas9-a0a247d711_lismn-2 2 16 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated in vitro In vitro 13 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) physical association physical association Extended Data Figure 1 cas9-a0a247d711_lismn-1 2 13 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 15 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) 16 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated in vitro In vitro physical association physical association Extended Data Figure 1 a0a247d711_lismn-cas9-3 2 13 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) 15 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified A0A247D711:p.Asn12Thr N12T mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain N T Tue Jun 30 17:34:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn12Thr' A0A247D711:p.Gly38Ala G38A mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain G A Tue Jun 30 19:05:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly38Ala' gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range A0A247D711:p.Asp23Arg D23R mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain D R Tue Jun 30 16:48:00 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp23Arg' ecoli Escherichia coli (strain K12) 16 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated in vitro In vitro physical association physical association Extended Data Figure 3b (AcrIIA4 mutant) a0a247d711_lismn-cas9-4 2 15 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified A0A247D711:p.Asn36Tyr N36Y mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N Y Mon Apr 29 11:10:17 BST 2019 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn36Tyr' Tue Jun 30 17:04:39 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn36Tyr' gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range A0A247D711:p.Asn39Arg N39R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N R Tue Jun 30 20:03:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn39Arg' A0A247D711:p.Glu70Arg E70R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E R Tue Jun 30 16:05:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu70Arg' A0A247D711:p.Asp69Arg D69R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D R Tue Jun 30 15:42:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp69Arg' A0A247D711:p.[Asp14Arg;Glu40Arg] D14R, E40R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E R certain certain sequence position certain certain certain sequence position certain D R Tue Jun 30 17:27:45 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asp14Arg;Glu40Arg]' A0A247D711:p.[Asn36Arg;Asp69Arg] N36R, D69R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N R certain certain sequence position certain certain certain sequence position certain D R Tue Jun 30 17:51:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Asn36Arg;Asp69Arg]' ecoli Escherichia coli (strain K12) 13 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) 16 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated in vitro In vitro physical association physical association Extended Data Figure 3b (AcrIIA4 mutant) a0a247d711_lismn-cas9-5 2 13 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey purified purified Q99ZW2:p.[Arg976Tyr;Lys1200Tyr] R976Y, K1200Y mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Y certain certain sequence position certain certain certain sequence position certain K Y Tue Jun 30 19:31:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Arg976Tyr;Lys1200Tyr]' Q99ZW2:p.[Lys1200Tyr;Ser1216Arg] K1200Y,S1216R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K Y certain certain sequence position certain certain certain sequence position certain S R Tue Jun 30 17:47:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys1200Tyr;Ser1216Arg]' Q99ZW2:p.Thr1138Tyr T1138Y mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain T Y Tue Jun 30 18:48:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr1138Tyr' Q99ZW2:p.Asp1332Arg D1332R mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D R Tue Jun 30 17:59:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp1332Arg' Q99ZW2:p.Arg1335Tyr R1335Y mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain R Y Tue Jun 30 17:27:27 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg1335Tyr' Q99ZW2:p.Ser1216Thr S1216T mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain S T Tue Jun 30 18:35:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser1216Thr' ecoli Escherichia coli (strain K12) 15 weight by comassie molecular weight estimation by coomasie staining weight by comassie experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait purified purified gst tag gst tag gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 16 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role ancillary ancillary in vitro translated in vitro translated protein in vitro translated in vitro In vitro physical association physical association Extended Data Figure 3b (SpyCas9 mutant) a0a247d711_lismn-cas9-6 1 16 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component in vitro translated in vitro translated protein in vitro translated in vitro In vitro 13 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) 15 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) physical association physical association Figure 4d