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/ khalil-2011-3 in vitro In vitro pull down pull down sequence tag sequence tag identification sequence tag Khalil S., Pawelek PD., Khalil S., Pawelek PD. Cells were iron starved. ppawelek@alcor.concordia.ca Biochemistry 2011 Only protein-protein interactions imex curation imex curation khalil-2011-1 in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca Biochemistry 2011 Only protein-protein interactions imex curation imex curation khalil-2011-2 in vitro In vitro dls dynamic light scattering dls predetermined predetermined participant predetermined Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca Biochemistry 2011 Only protein-protein interactions imex curation imex curation khalil-2011-5 in vitro In vitro fret fluorescent resonance energy transfer FRET RET FRET analysis FRET RET FRET analysis fret predetermined predetermined participant predetermined Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca Biochemistry 2011 Only protein-protein interactions imex curation imex curation khalil-2011-4 in vitro In vitro crosslink cross-linking study crosslink sequence tag sequence tag identification sequence tag Khalil S., Pawelek PD., Khalil S., Pawelek PD. Crosslinker SMCC (succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate) ppawelek@alcor.concordia.ca Biochemistry 2011 Only protein-protein interactions imex curation imex curation khalil-2011-6 in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps predetermined predetermined participant predetermined Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca Biochemistry 2011 Only protein-protein interactions imex curation imex curation khalil-2011-7 in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca Biochemistry 2011 Only protein-protein interactions imex curation imex curation ente_ecoli Enterobactin synthase component E entE Enterochelin synthase E 2,3-dihydroxybenzoate-AMP synthase Dihydroxybenzoic acid-activating enzyme S-dihydroxybenzoyltransferase JW0586 b0594 2,3-dihydroxybenzoate-AMP ligase protein protein ecoli Escherichia coli (strain K12) MSIPFTRWPEEFARRYREKGYWQDLPLTDILTRHAASDSIAVIDGERQLSYRELNQAADNLACSLRRQGIKPGETALVQLGNVAELYITFFALLKLGVAPVLALFSHQRSELNAYASQIEPALLIADRQHALFSGDDFLNTFVTEHSSIRVVQLLNDSGEHNLQDAINHPAEDFTATPSPADEVAYFQLSGGTTGTPKLIPRTHNDYYYSVRRSVEICQFTQQTRYLCAIPAAHNYAMSSPGSLGVFLAGGTVVLAADPSATLCFPLIEKHQVNVTALVPPAVSLWLQALIEGESRAQLASLKLLQVGGARLSATLAARIPAEIGCQLQQVFGMAEGLVNYTRLDDSAEKIIHTQGYPMCPDDEVWVADAEGNPLPQGEVGRLMTRGPYTFRGYYKSPQHNASAFDANGFYCSGDLISIDPEGYITVQGREKDQINRGGEKIAAEEIENLLLRHPAVIYAALVSMEDELMGEKSCAYLVVKEPLRAVQVRRFLREQGIAEFKLPDRVECVDSLPLTAVGKVDKKQLRQWLASRASA enta_ecoli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase entA Trans-2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase JW0588 b0596 protein protein ecoli Escherichia coli (strain K12) MDFSGKNVWVTGAGKGIGYATALAFVEAGAKVTGFDQAFTQEQYPFATEVMDVADAAQVAQVCQRLLAETERLDALVNAAGILRMGATDQLSKEDWQQTFAVNVGGAFNLFQQTMNQFRRQRGGAIVTVASDAAHTPRIGMSAYGASKAALKSLALSVGLELAGSGVRCNVVSPGSTDTDMQRTLWVSDDAEEQRIRGFGEQFKLGIPLGKIARPQEIANTILFLASDLASHITLQDIVVDGGSTLGA entb_ecoli Enterobactin synthase component B entB entG b0595 Enterochelin synthase B JW0587 Enterobactin biosynthesis bifunctional protein EntB protein protein ecoli Escherichia coli (strain K12) MAIPKLQAYALPESHDIPQNKVDWAFEPQRAALLIHDMQDYFVSFWGENCPMMEQVIANIAALRDYCKQHNIPVYYTAQPKEQSDEDRALLNDMWGPGLTRSPEQQKVVDRLTPDADDTVLVKWRYSAFHRSPLEQMLKESGRNQLIITGVYAHIGCMTTATDAFMRDIKPFMVADALADFSRDEHLMSLKYVAGRSGRVVMTEELLPAPIPASKAALREVILPLLDESDEPFDDDNLIDYGLDSVRMMALAARWRKVHGDIDFVMLAKNPTIDAWWKLLSREVK 2,3-dihydroxybenzoic acid 2,3-dihydroxybenzoic acid 2,3 DHB 2,3-DIHYDROXY-BENZOIC ACID 2,3-dihydroxybenzoic acid 2,3-Dihydroxybenzoic acid 2-pyrocatechuic acid 3-hydroxysalicylic acid catechol-3-carboxylic acid DOBK o-pyrocatechuic acid pyrocatechuic acid 2,3-dihydroxybenzoic acid small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) enta-ente-1 3710248 N/A 3710255 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level ecoli Escherichia coli (strain K12) N/A 3710256 unspecified role unspecified role bait bait purified purified n-terminal 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 ecoli Escherichia coli (strain K12) physical association physical association false false false Fig.2A, Supp. Table 1 Interaction observed in the presence or absence of 2,3-dihydroxybenzoate (DHB) enta-ente-2 3710248 N/A 3710256 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level ecoli Escherichia coli (strain K12) N/A 3710255 unspecified role unspecified role bait bait purified purified n-terminal 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 ecoli Escherichia coli (strain K12) physical association physical association false false false Fig.2B, Supp. Table 1 Interaction observed in the absence or absence of 2,3-dihydroxybenzoate (DHB) ente-enta-1 3710248 N/A 3710257 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level ecoli Escherichia coli (strain K12) N/A 3710256 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level ecoli Escherichia coli (strain K12) N/A 3710255 unspecified role unspecified role bait bait purified purified n-terminal 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 ecoli Escherichia coli (strain K12) association association false false false Fig.2B, Supp. Table 1 Interaction observed in the presence of 2,3-dihydroxybenzoate (DHB) enta-1 3710249 N/A 3710256 unspecified role unspecified role neutral component neutral component purified purified n-terminal 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 ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Fig.1 Table 1 enta-2 3710249 N/A 3710256 unspecified role unspecified role neutral component neutral component purified purified n-terminal 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 ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 direct interaction direct interaction false false false Fig.1 Table 1 enta-3 3710250 N/A 3710256 unspecified role unspecified role neutral component neutral component purified purified n-terminal 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 ecoli Escherichia coli (strain K12) Stoichiometry: 2.0 direct interaction direct interaction false false false Table 2 enta-4 3710250 N/A 3710256 unspecified role unspecified role neutral component neutral component purified purified n-terminal 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 ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 direct interaction direct interaction false false false Table 2 2,3_dihydroxybenzoic acid-ente-1 3710251 N/A 3710258 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept N/A 3710255 unspecified role unspecified role fluorescence donor fluorescence donor purified purified n-terminal 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig.4,6 2,3_dihydroxybenzoic acid-enta-1 3710251 N/A 3710256 competitor competitor competitor unspecified role unspecified role purified purified n-terminal 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 ecoli Escherichia coli (strain K12) N/A 3710258 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept N/A 3710255 unspecified role unspecified role fluorescence donor fluorescence donor purified purified n-terminal 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Figs.4, 6 enta-ente-3 3710252 N/A 3710256 unspecified role unspecified role neutral component neutral component purified purified n-terminal 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 ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 N/A 3710255 unspecified role unspecified role neutral component neutral component purified purified n-terminal 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 ecoli Escherichia coli (strain K12) Stoichiometry: 4.0 direct interaction direct interaction false false false Supp. Table 2, Fig. S1 2,3_dihydroxybenzoic acid-ente-2 3710253 N/A 3710258 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept N/A 3710255 unspecified role unspecified role neutral component neutral component purified purified n-terminal 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig.5 No additional increase in anisotropy was observed with the addition of entA ente-enta-2 3710254 N/A 3710256 unspecified role unspecified role neutral component neutral component purified purified n-terminal 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 ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 N/A 3710255 unspecified role unspecified role neutral component neutral component purified purified n-terminal 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 ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false Fig.7, Table 3