IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ jenkins-2006-1 Human C4b-binding protein, structural basis for interaction with streptococcal M protein, a major bacterial virulence factor. 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 J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation jenkins-2006-2 Human C4b-binding protein, structural basis for interaction with streptococcal M protein, a major bacterial virulence factor. 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 J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation jenkins-2006-3 Human C4b-binding protein, structural basis for interaction with streptococcal M protein, a major bacterial virulence factor. 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 J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation arp4_strpy IgA receptor arp4 protein protein strpy Streptococcus pyogenes MARKDTNKQYSLRKLKTGTASVAVAVAVLGAGFANQTEVKAAEIKKPQADSAWNWPKEYNALLKENEELKVEREKYLSYADDKEKDPQYRALMGENQDLRKREGQYQDKIEELEKERKEKQERQEQLERQYQIEADKHYQEQQKKHQQEQQQLEAEKQKLAKDKQISDASRQGLSRDLEASRAAKKELEAEHQKLKEEKQISDASRQGLSRDLEASREAKKKVEADLAALTAEHQKLKEDKQISDASRQGLSRDLEASREAKKKVEADLAEANSKLQALEKLNKELEEGKKLSEKEKAELQARLEAEAKALKEQLAKQAEELAKLKGNQTPNAKVAPQANRSRSAMTQQKRTLPSTGETANPFFTAAAATVMVSAGMLALKRKEEN c4bpa_human C4b-binding protein alpha chain Proline-rich protein C4BPA C4BP protein protein human Homo sapiens Human MHPPKTPSGALHRKRKMAAWPFSRLWKVSDPILFQMTLIAALLPAVLGNCGPPPTLSFAAPMDITLTETRFKTGTTLKYTCLPGYVRSHSTQTLTCNSDGEWVYNTFCIYKRCRHPGELRNGQVEIKTDLSFGSQIEFSCSEGFFLIGSTTSRCEVQDRGVGWSHPLPQCEIVKCKPPPDIRNGRHSGEENFYAYGFSVTYSCDPRFSLLGHASISCTVENETIGVWRPSPPTCEKITCRKPDVSHGEMVSGFGPIYNYKDTIVFKCQKGFVLRGSSVIHCDADSKWNPSPPACEPNSCINLPDIPHASWETYPRPTKEDVYVVGTVLRYRCHPGYKPTTDEPTTVICQKNLRWTPYQGCEALCCPEPKLNNGEITQHRKSRPANHCVYFYGDEISFSCHETSRFSAICQGDGTWSPRTPSCGDICNFPPKIAHGHYKQSSSYSFFKEEIIYECDKGYILVGQAKLSCSYSHWSAPAPQCKALCRKPELVNGRLSVDKDQYVEPENVTIQCDSGYGVVGPQSITCSGNRTWYPEVPKCEWETPEGCEQVLTGKRLMQCLPNPEDVKMALEVYKLSLEIEQLELQRDSARQSTLDKEL apr4-c4bp Binding of C4BP and APR4 501868 N/A 501871 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) N/A 501872 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 false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 1A apr4-c4bp-1 Binding of C4BP and APR4 501868 N/A 501872 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 false ecoli Escherichia coli (strain K12) N/A 501871 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 1A apr4-c4bp-2 Binding of C4BP and APR4 501868 N/A 501872 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) N/A 501871 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 1B Insertion of two alanine residues between Lysine 188 and Isoleucine 189 resulted in abolished interaction between the two proteins. apr4-c4bp-3 Binding of C4BP and APR4 501869 N/A 501872 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 false n terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 501871 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 false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false Figure 1C 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 Binding of C4BP and APR4 demonstrated by NMR 501870 N/A 501872 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 501871 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 false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false Figure 4 and Figure 5 The APR4 protein was present as a dimer.