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/ Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Biochemistry 2011 imex curation Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca 13-Nov-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-OCT-13 AT 07:54 BST AT 07:54 BST by LIVIAP Biochemistry 2011 Only protein-protein interactions imex curation imex curation Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Biochemistry 2011 imex curation Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca 13-Nov-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-OCT-13 AT 07:54 BST AT 07:54 BST by LIVIAP Biochemistry 2011 Only protein-protein interactions imex curation imex curation Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Biochemistry 2011 imex curation Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca 13-Nov-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-OCT-13 AT 07:54 BST AT 07:54 BST by LIVIAP Biochemistry 2011 Only protein-protein interactions imex curation imex curation Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Biochemistry 2011 imex curation Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca 13-Nov-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-OCT-13 AT 07:54 BST AT 07:54 BST by LIVIAP Biochemistry 2011 Only protein-protein interactions imex curation imex curation Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Biochemistry 2011 imex curation Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca 13-Nov-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-OCT-13 AT 07:54 BST AT 07:54 BST by LIVIAP Biochemistry 2011 Only protein-protein interactions imex curation imex curation Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Biochemistry 2011 imex curation Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca 13-Nov-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-OCT-13 AT 07:54 BST AT 07:54 BST by LIVIAP Biochemistry 2011 Only protein-protein interactions imex curation imex curation Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Enzymatic adenylation of 2,3-dihydroxybenzoate is enhanced by a protein-protein interaction between Escherichia coli 2,3-dihydro-2,3-dihydroxybenzoate dehydrogenase (EntA) and 2,3-dihydroxybenzoate-AMP ligase (EntE). Biochemistry 2011 imex curation Khalil S., Pawelek PD., Khalil S., Pawelek PD. ppawelek@alcor.concordia.ca 13-Nov-2020: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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 Accepted 2020-OCT-13 AT 07:54 BST AT 07:54 BST by LIVIAP Biochemistry 2011 Only protein-protein interactions imex curation imex curation 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) 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 DIP protein P0ADI4 original sequence version: null E98C62D5ED23BAB1 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 DIP protein P10378 original sequence version: null F818942DFDD8DC99 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 E840488335AD317B enta-1 7 11 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.1 Table 1 enta-2 7 11 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.1 Table 1 enta-3 5 11 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Table 2 enta-4 5 11 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Table 2 enta-ente-1 6 10 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) 11 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 ecoli Escherichia coli (strain K12) physical association physical association Fig.2A, Supp. Table 1 Interaction observed in the presence or absence of 2,3-dihydroxybenzoate (DHB) enta-ente-2 6 11 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) 10 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 ecoli Escherichia coli (strain K12) physical association physical association Fig.2B, Supp. Table 1 Interaction observed in the absence or absence of 2,3-dihydroxybenzoate (DHB) ente-enta-1 6 9 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level ecoli Escherichia coli (strain K12) 11 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) 10 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 ecoli Escherichia coli (strain K12) association association Fig.2B, Supp. Table 1 Interaction observed in the presence of 2,3-dihydroxybenzoate (DHB) ente-enta-2 1 11 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 ecoli Escherichia coli (strain K12) 10 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.7, Table 3 2,3_dihydroxybenzoic acid-ente-1 4 8 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept 10 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.4,6 2,3_dihydroxybenzoic acid-enta-1 4 11 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 ecoli Escherichia coli (strain K12) 8 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept 10 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Figs.4, 6 2,3_dihydroxybenzoic acid-ente-2 2 8 unspecified role unspecified role fluorescence accept fluorescence acceptor fluorescence accept 10 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig.5 No additional increase in anisotropy was observed with the addition of entA enta-ente-3 3 11 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 ecoli Escherichia coli (strain K12) 10 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Supp. Table 2, Fig. S1