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/ kim-2011f-1 Crystal structure of β-barrel assembly machinery BamCD protein complex. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by comassie molecular weight estimation by coomasie staining weight by comassie Kim K.H., Aulakh S., Paetzel M. kkim@sfu.ca, ska16@sfu.ca, mpaetzel@sfu.ca J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation kim-2011f-2 Crystal structure of β-barrel assembly machinery BamCD protein complex. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Kim K.H., Aulakh S., Paetzel M. kkim@sfu.ca, ska16@sfu.ca, mpaetzel@sfu.ca J. Biol. Chem. (0021-9258) 2011 Only protein-protein interactions imex curation bamc_ecoli Outer membrane protein assembly factor BamC bamC dapX nlpB JW2462 b2477 protein protein ecoli Escherichia coli (strain K12) MAYSVQKSRLAKVAGVSLVLLLAACSSDSRYKRQVSGDEAYLEAAPLAELHAPAGMILPVTSGDYAIPVTNGSGAVGKALDIRPPAQPLALVSGARTQFTGDTASLLVENGRGNTLWPQVVSVLQAKNYTITQRDDAGQTLTTDWVQWNRLDEDEQYRGRYQISVKPQGYQQAVTVKLLNLEQAGKPVADAASMQRYSTEMMNVISAGLDKSATDAANAAQNRASTTMDVQSAADDTGLPMLVVRGPFNVVWQRLPAALEKVGMKVTDSTRSQGNMAVTYKPLSDSDWQELGASDPGLASGDYKLQVGDLDNRSSLQFIDPKGHTLTQSQNDALVAVFQAAFSK bamd_ecoli Outer membrane protein assembly factor BamD bamD yfiO JW2577 b2595 protein protein ecoli Escherichia coli (strain K12) MTRMKYLVAAATLSLFLAGCSGSKEEVPDNPPNEIYATAQQKLQDGNWRQAITQLEALDNRYPFGPYSQQVQLDLIYAYYKNADLPLAQAAIDRFIRLNPTHPNIDYVMYMRGLTNMALDDSALQGFFGVDRSDRDPQHARAAFSDFSKLVRGYPNSQYTTDATKRLVFLKDRLAKYEYSVAEYYTERGAWVAVVNRVEGMLRDYPDTQATRDALPLMENAYRQMQMNAQAEKVAKIIAANSSNT bamc-bamd 1306883 n/a 1306885 biological role biological role neutral component neutral component 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 false region sufficient to bind sufficient binding region sufficient to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1306886 biological role biological role neutral component neutral component 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 false region sufficient to bind sufficient binding region sufficient to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1306893 1306890 direct interaction direct interaction false false false FIGURE 1. Formation of the BamCD complex. bamc-bamd-1 1306883 n/a 1306886 biological role biological role neutral component neutral component 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 false region sufficient to bind sufficient binding region sufficient to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1306885 biological role biological role neutral component neutral component region required to bind necessary binding region required to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain false tag 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 ecoli Escherichia coli (strain K12) 1306899 1306897 direct interaction direct interaction false false false FIGURE 1. Formation of the BamCD complex bamc-bamd-2 1306884 n/a 1306886 biological role biological role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1306885 biological role biological role neutral component neutral component region required to bind necessary binding region required to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1306905 1306903 direct interaction direct interaction false false false Figure 2, Figure 3 Although the full-length BamCD complex was used for crystallization, BamC was cleaved to a smaller fragment corresponding to BamCUN (Asp28–Ala217) in the crystallization drop during incubation. BamC has been previously shown to be susceptible to degradation in the linker region connecting the N- and C-terminal domains. 2.9A R-Value: 0.181 (obs.), R-Free: 0.243