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/ ahuja-2005-1 CcmD is involved in complex formation between CcmC and the heme chaperone CcmE during cytochrome c maturation. ecoli Escherichia coli (strain K12) anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Ahuja U., Thony-Meyer L. Location of Strep tag was found in Schultz et al,. 2002. lthoeny@micro.biol.ethz.ch J. Biol. Chem. (0021-9258) 2005 Only protein-protein interactions imex curation ahuja-2005-2 CcmD is involved in complex formation between CcmC and the heme chaperone CcmE during cytochrome c maturation. ecoli Escherichia coli (strain K12) anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Ahuja U., Thony-Meyer L. Location of Strep tag was found in Schultz et al,. 2002. lthoeny@micro.biol.ethz.ch J. Biol. Chem. (0021-9258) 2005 Only protein-protein interactions imex curation ahuja-2005-3 CcmD is involved in complex formation between CcmC and the heme chaperone CcmE during cytochrome c maturation. ecoli Escherichia coli (strain K12) pull down pull down anti tag western anti tag western blot anti tag western Ahuja U., Thony-Meyer L. StrepTactin-Sepharose affinity chromatography Location of Strep tag was found in Schultz et al,. 2002. lthoeny@micro.biol.ethz.ch J. Biol. Chem. (0021-9258) 2005 Only protein-protein interactions imex curation ahuja-2005-4 CcmD is involved in complex formation between CcmC and the heme chaperone CcmE during cytochrome c maturation. ecoli Escherichia coli (strain K12) pull down pull down western blot western blot Immuno blot Immuno blot Ahuja U., Thony-Meyer L. nickel affinity chromatography Location of Strep tag was found in Schultz et al,. 2002. lthoeny@micro.biol.ethz.ch J. Biol. Chem. (0021-9258) 2005 Only protein-protein interactions imex curation ccmc_ecoli Heme exporter protein C ccmC yejT yejU Cytochrome c-type biogenesis protein CcmC b2199 JW2187 protein protein ecoli Escherichia coli (strain K12) MWKTLHQLAIPPRLYQICGWFIPWLAIASVVVLTVGWIWGFGFAPADYQQGNSYRIIYLHVPAAIWSMGIYASMAVAAFIGLVWQMKMANLAVAAMAPIGAVFTFIALVTGSAWGKPMWGTWWVWDARLTSELVLLFLYVGVIALWHAFDDRRLAGRAAGILVLIGVVNLPIIHYSVEWWNTLHQGSTRMQQSIDPAMRSPLRWSIFGFLLLSATLTLMRMRNLILLMEKRRPWVSELILKRGRK ccmd_ecoli Heme exporter protein D ccmD yojM Cytochrome c-type biogenesis protein CcmD b2198 JW2186 protein protein ecoli Escherichia coli (strain K12) MTPAFASWNEFFAMGGYAFFVWLAVVMTVIPLVVLVVHSVMQHRAILRGVAQQRAREARLRAAQQQEAA ccme_ecoli Cytochrome c-type biogenesis protein CcmE ccmE b2197 JW2185 yejS Cytochrome c maturation protein E Heme chaperone CcmE protein protein ecoli Escherichia coli (strain K12) MNIRRKNRLWIACAVLAGLALTIGLVLYALRSNIDLFYTPGEILYGKRETQQMPEVGQRLRVGGMVMPGSVQRDPNSLKVTFTIYDAEGSVDVSYEGILPDLFREGQGVVVQGELEKGNHILAKEVLAKHDENYTPPEVEKAMEANHRRPASVYKDPAS ccmc-ccmd 1144290 n/a 1144294 unspecified role unspecified role bait bait c_terminus 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/a 1144295 unspecified role unspecified role prey prey c_terminus strep ii tag strep ii tag Strep (II) Strep II Strep (II) Strep II c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig 3B ccme-ccmd 1144291 n/a 1144295 unspecified role unspecified role prey prey c_terminus strep ii tag strep ii tag Strep (II) Strep II Strep (II) Strep II c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 1144296 unspecified role unspecified role bait bait physical association physical association false false false Fig 3B ccmd-ccmc 1144292 n/a 1144294 unspecified role unspecified role prey prey c_terminus 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/a 1144295 unspecified role unspecified role bait bait c_terminus strep ii tag strep ii tag Strep (II) Strep II Strep (II) Strep II c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig 4A ccmc-ccme-2 1144293 n/a 1144294 unspecified role unspecified role bait bait c_terminus 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/a 1144296 unspecified role unspecified role prey prey n/a 1144295 unspecified role unspecified role ancillary ancillary physical association physical association false false false Fig 5 CcmE co-eluted with CcmC only in the presence of CcmD (Fig. 5A), but not in its absence.