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/ lin-2009c-1 MacB ABC transporter is a dimer whose ATPase activity and macrolide-binding capacity are regulated by the membrane fusion protein MacA. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Lin H.T., Bavro V.N., Barrera N.P., Frankish H.M., Velamakanni S., van Veen H.W., Robinson C.V., Borges-Walmsley M.I., Walmsley A.R. a.r.walmsley@durham.ac.uk. J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation lin-2009c-7 MacB ABC transporter is a dimer whose ATPase activity and macrolide-binding capacity are regulated by the membrane fusion protein MacA. in vitro In vitro atpase assay atpase assay predetermined predetermined participant predetermined Lin H.T., Bavro V.N., Barrera N.P., Frankish H.M., Velamakanni S., van Veen H.W., Robinson C.V., Borges-Walmsley M.I., Walmsley A.R. a.r.walmsley@durham.ac.uk. J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation lin-2009c-2 MacB ABC transporter is a dimer whose ATPase activity and macrolide-binding capacity are regulated by the membrane fusion protein MacA. in vitro In vitro crosslink cross-linking study crosslink sequence tag sequence tag identification sequence tag Lin H.T., Bavro V.N., Barrera N.P., Frankish H.M., Velamakanni S., van Veen H.W., Robinson C.V., Borges-Walmsley M.I., Walmsley A.R. a.r.walmsley@durham.ac.uk. J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation lin-2009c-3 MacB ABC transporter is a dimer whose ATPase activity and macrolide-binding capacity are regulated by the membrane fusion protein MacA. in vitro In vitro crosslink cross-linking study crosslink predetermined predetermined participant predetermined Lin H.T., Bavro V.N., Barrera N.P., Frankish H.M., Velamakanni S., van Veen H.W., Robinson C.V., Borges-Walmsley M.I., Walmsley A.R. a.r.walmsley@durham.ac.uk. J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation lin-2009c-4 MacB ABC transporter is a dimer whose ATPase activity and macrolide-binding capacity are regulated by the membrane fusion protein MacA. ecoli Escherichia coli (strain K12) cosedimentation cosedimentation weight identificat confirmation by molecular weight weight identificat Lin H.T., Bavro V.N., Barrera N.P., Frankish H.M., Velamakanni S., van Veen H.W., Robinson C.V., Borges-Walmsley M.I., Walmsley A.R. a.r.walmsley@durham.ac.uk. J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation lin-2009c-5 MacB ABC transporter is a dimer whose ATPase activity and macrolide-binding capacity are regulated by the membrane fusion protein MacA. in vitro In vitro atomic force microsc atomic force microscopy AFM AFM atomic force microsc predetermined predetermined participant predetermined Lin H.T., Bavro V.N., Barrera N.P., Frankish H.M., Velamakanni S., van Veen H.W., Robinson C.V., Borges-Walmsley M.I., Walmsley A.R. a.r.walmsley@durham.ac.uk. J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation lin-2009c-6 MacB ABC transporter is a dimer whose ATPase activity and macrolide-binding capacity are regulated by the membrane fusion protein MacA. ecoli Escherichia coli (strain K12) ms of complexes mass spectrometry studies of complexes ms of complexes predetermined predetermined participant predetermined Lin H.T., Bavro V.N., Barrera N.P., Frankish H.M., Velamakanni S., van Veen H.W., Robinson C.V., Borges-Walmsley M.I., Walmsley A.R. a.r.walmsley@durham.ac.uk. J. Biol. Chem. (0021-9258) 2009 Only protein-protein interactions imex curation maca_ecoli Macrolide export protein MacA macA b0878 ybjY JW0862 protein protein ecoli Escherichia coli (strain K12) MKKRKTVKKRYVIALVIVIAGLITLWRILNAPVPTYQTLIVRPGDLQQSVLATGKLDALRKVDVGAQVSGQLKTLSVAIGDKVKKDQLLGVIDPEQAENQIKEVEATLMELRAQRQQAEAELKLARVTYSRQQRLAQTKAVSQQDLDTAATEMAVKQAQIGTIDAQIKRNQASLDTAKTNLDYTRIVAPMAGEVTQITTLQGQTVIAAQQAPNILTLADMSAMLVKAQVSEADVIHLKPGQKAWFTVLGDPLTRYEGQIKDVLPTPEKVNDAIFYYARFEVPNPNGLLRLDMTAQVHIQLTDVKNVLTIPLSALGDPVGDNRYKVKLLRNGETREREVTIGARNDTDVEIVKGLEAGDEVVIGEAKPGAAQ tolc_ecoli Outer membrane protein TolC tolC mtcB mukA refI Multidrug efflux pump subunit TolC Outer membrane factor TolC colE1-i toc weeA JW5503 b3035 protein protein ecoli Escherichia coli (strain K12) MKKLLPILIGLSLSGFSSLSQAENLMQVYQQARLSNPELRKSAADRDAAFEKINEARSPLLPQLGLGADYTYSNGYRDANGINSNATSASLQLTQSIFDMSKWRALTLQEKAAGIQDVTYQTDQQTLILNTATAYFNVLNAIDVLSYTQAQKEAIYRQLDQTTQRFNVGLVAITDVQNARAQYDTVLANEVTARNNLDNAVEQLRQITGNYYPELAALNVENFKTDKPQPVNALLKEAEKRNLSLLQARLSQDLAREQIRQAQDGHLPTLDLTASTGISDTSYSGSKTRGAAGTQYDDSNMGQNKVGLSFSLPIYQGGMVNSQVKQAQYNFVGASEQLESAHRSVVQTVRSSFNNINASISSINAYKQAVVSAQSSLDAMEAGYSVGTRTIVDVLDATTTLYNAKQELANARYNYLINQLNIKSALGTLNEQDLLALNNALSKPVSTNPENVAPQTPEQNAIADGYAPDSPAPVVQQTSARTTTSNGHNPFRN macb_ecoli Macrolide export ATP-binding/permease protein MacB macB b0879 JW0863 ybjZ protein protein ecoli Escherichia coli (strain K12) MTPLLELKDIRRSYPAGDEQVEVLKGISLDIYAGEMVAIVGASGSGKSTLMNILGCLDKATSGTYRVAGQDVATLDADALAQLRREHFGFIFQRYHLLSHLTAEQNVEVPAVYAGLERKQRLLRAQELLQRLGLEDRTEYYPAQLSGGQQQRVSIARALMNGGQVILADEPTGALDSHSGEEVMAILHQLRDRGHTVIIVTHDPQVAAQAERVIEIRDGEIVRNPPAIEKVNVTGGTEPVVNTVSGWRQFVSGFNEALTMAWRALAANKMRTLLTMLGIIIGIASVVSIVVVGDAAKQMVLADIRSIGTNTIDVYPGKDFGDDDPQYQQALKYDDLIAIQKQPWVASATPAVSQNLRLRYNNVDVAASANGVSGDYFNVYGMTFSEGNTFNQEQLNGRAQVVVLDSNTRRQLFPHKADVVGEVILVGNMPARVIGVAEEKQSMFGSSKVLRVWLPYSTMSGRVMGQSWLNSITVRVKEGFDSAEAEQQLTRLLSLRHGKKDFFTWNMDGVLKTVEKTTRTLQLFLTLVAVISLVVGGIGVMNIMLVSVTERTREIGIRMAVGARASDVLQQFLIEAVLVCLVGGALGITLSLLIAFTLQLFLPGWEIGFSPLALLLAFLCSTVTGILFGWLPARNAARLDPVDALARE atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) tolc-maca 1318837 n/a 1318844 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation stag s tag s tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1318845 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false Figure 1 macb-tolc 1318837 n/a 1318845 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation stag s tag s tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1318846 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) physical association physical association false false false Figure 1 They detected interaction between TolC and MacB but the intensity of the band suggested a weak interaction macb-maca-1 1318837 n/a 1318846 unspecified role unspecified role bait bait nucl transformation nucleic acid transformation nucl transformation histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1318844 unspecified role unspecified role prey prey nucl transformation nucleic acid transformation nucl transformation s tag tag tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false 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) physical association physical association false false false Figure 1 atp-macb 1318838 n/a 1318846 unspecified role unspecified role unspecified role unspecified role histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1318847 enzyme target enzyme target substrate substrate unspecified role unspecified role direct interaction direct interaction false false false Figure 5 atp-maca 1318838 n/a 1318847 enzyme target enzyme target substrate substrate unspecified role unspecified role n/a 1318846 enzyme enzyme unspecified role unspecified role histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1318844 inhibitor inhibitor unspecified role unspecified role histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figure 5 maca-macb-1 1318839 n/a 1318846 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1318844 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Supplemental data Figure 2A crosslinking agent was DMS and participant detection method SDS-PAGE and MALDI-TOF. macb-1 1318840 n/a 1318846 unspecified role unspecified role neutral component neutral component purified purified histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Supplemental data Figure 2B The most abundant band on the gel was indicative of a dimer. However, there were several higher-Mr bands also present that would suggest that the protein can form higher-order oligomers. macb-1 1318841 n/a 1318846 unspecified role unspecified role neutral component neutral component nucl transformation nucleic acid transformation nucl transformation histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Figure 3A, 3B macb-1 1318842 n/a 1318846 unspecified role unspecified role neutral component neutral component purified purified histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Figure 4 macb-1 1318843 n/a 1318846 unspecified role unspecified role unspecified role unspecified role histag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Figure 3C