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/ Structural determinants for the binding of anthrax lethal factor to oligomeric protective antigen. Structural determinants for the binding of anthrax lethal factor to oligomeric protective antigen. J. Biol. Chem. (0021-9258) 2006 imex curation Melnyk RA., Hewitt KM., Lacy DB., Lin HC., Gessner CR., Li S., Woods VL., Collier RJ. Hydrogen-deuterium exchange MS was performed. The number of deuterons incorporated into proteins was determined by MS of peptides obtained by passing protein through an immobilised protease column. vwoods@ucsd.edu,jcollier@hms.harvard.edu 13-SEP-2006: Contacted by JYOTI Only protein-protein interactions in vitro In vitro ms of complexes mass spectrometry studies of complexes ms of complexes predetermined predetermined participant predetermined Melnyk RA., Hewitt KM., Lacy DB., Lin HC., Gessner CR., Li S., Woods VL., Collier RJ. Hydrogen-deuterium exchange MS was performed. The number of deuterons incorporated into proteins was determined by MS of peptides obtained by passing protein through an immobilised protease column. Mutation mapping shows that the "mature" protein - minus the signal sequence was used. vwoods@ucsd.edu,jcollier@hms.harvard.edu Accepted 2006-AUG-29 by JYOTI. 13-SEP-2006: Contacted by JYOTI J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation lef_bacan Lethal factor Anthrax lethal toxin endopeptidase component lef pXO1-107 BXA0172 GBAA_pXO1_0172 protein protein bacan Bacillus anthracis MNIKKEFIKVISMSCLVTAITLSGPVFIPLVQGAGGHGDVGMHVKEKEKNKDENKRKDEERNKTQEEHLKEIMKHIVKIEVKGEEAVKKEAAEKLLEKVPSDVLEMYKAIGGKIYIVDGDITKHISLEALSEDKKKIKDIYGKDALLHEHYVYAKEGYEPVLVIQSSEDYVENTEKALNVYYEIGKILSRDILSKINQPYQKFLDVLNTIKNASDSDGQDLLFTNQLKEHPTDFSVEFLEQNSNEVQEVFAKAFAYYIEPQHRDVLQLYAPEAFNYMDKFNEQEINLSLEELKDQRMLARYEKWEKIKQHYQHWSDSLSEEGRGLLKKLQIPIEPKKDDIIHSLSQEEKELLKRIQIDSSDFLSTEEKEFLKKLQIDIRDSLSEEEKELLNRIQVDSSNPLSEKEKEFLKKLKLDIQPYDINQRLQDTGGLIDSPSINLDVRKQYKRDIQNIDALLHQSIGSTLYNKIYLYENMNINNLTATLGADLVDSTDNTKINRGIFNEFKKNFKYSISSNYMIVDINERPALDNERLKWRIQLSPDTRAGYLENGKLILQRNIGLEIKDVQIIKQSEKEYIRIDAKVVPKSKIDTKIQEAQLNINQEWNKALGLPKYTKLITFNVHNRYASNIVESAYLILNEWKNNIQSDLIKKVTNYLVDGNGRFVFTDITLPNIAEQYTHQDEIYEQVHSKGLYVPESRSILLHGPSKGVELRNDSEGFIHEFGHAVDDYAGYLLDKNQSDLVTNSKKFIDIFKEEGSNLTSYGRTNEAEFFAEAFRLMHSTDHAERLKVQKNAPKTFQFINDQIKFIINS DIP protein P15917 original sequence version: 109 DIP protein P15917 original sequence version: 138 DIP protein P15917 original sequence version: 93 DIP protein P15917 original sequence version: 98 DIP protein P15917 original sequence version: 112 DIP protein P15917 original sequence version: 125 2076B4D7277317EE pag_bacan Protective antigen PA-83 Anthrax toxins translocating protein pagA pag pXO1-110 BXA0164 GBAA_pXO1_0164 protein protein bacan Bacillus anthracis MKKRKVLIPLMALSTILVSSTGNLEVIQAEVKQENRLLNESESSSQGLLGYYFSDLNFQAPMVVTSSTTGDLSIPSSELENIPSENQYFQSAIWSGFIKVKKSDEYTFATSADNHVTMWVDDQEVINKASNSNKIRLEKGRLYQIKIQYQRENPTEKGLDFKLYWTDSQNKKEVISSDNLQLPELKQKSSNSRKKRSTSAGPTVPDRDNDGIPDSLEVEGYTVDVKNKRTFLSPWISNIHEKKGLTKYKSSPEKWSTASDPYSDFEKVTGRIDKNVSPEARHPLVAAYPIVHVDMENIILSKNEDQSTQNTDSQTRTISKNTSTSRTHTSEVHGNAEVHASFFDIGGSVSAGFSNSNSSTVAIDHSLSLAGERTWAETMGLNTADTARLNANIRYVNTGTAPIYNVLPTTSLVLGKNQTLATIKAKENQLSQILAPNNYYPSKNLAPIALNAQDDFSSTPITMNYNQFLELEKTKQLRLDTDQVYGNIATYNFENGRVRVDTGSNWSEVLPQIQETTARIIFNGKDLNLVERRIAAVNPSDPLETTKPDMTLKEALKIAFGFNEPNGNLQYQGKDITEFDFNFDQQTSQNIKNQLAELNATNIYTVLDKIKLNAKMNILIRDKRFHYDRNNIAVGADESVVKEAHREVINSSTEGLLLNIDKDIRKILSGYIVEIEDTEGLKEVINDRYDMLNISSLRQDGKTFIDFKKYNDKLPLYISNPNYKVNVYAVTKENTIINPSENGDTSTNGIKKILIFSKKGYEIG DIP protein P13423 original sequence version: 106 DIP protein P13423 original sequence version: 91 DIP protein P13423 original sequence version: 93 DIP protein P13423 original sequence version: 110 DIP protein P13423 original sequence version: 124 DIP protein P13423 original sequence version: 150 3AE1EFBF48FAA03F lef-paga 1 3 unspecified role unspecified role neutral component neutral component c-terminal PA83 sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain range range P13423:p.Asp541Lys mutation mutation certain certain sequence position certain certain certain sequence position certain D K Wed May 02 17:44:00 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp541lys' Tue Jun 30 15:36:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp541Lys' P13423:p.[Lys228Glu;Arg497Ala;Arg499Asp] mutation mutation certain certain sequence position certain certain certain sequence position certain K E certain certain sequence position certain certain certain sequence position certain R A certain certain sequence position certain certain certain sequence position certain R D Wed May 02 17:26:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys228glu,arg497ala,arg499asp' Tue Jun 30 17:21:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Lys228Glu;Arg497Ala;Arg499Asp]' ecoli Escherichia coli (strain K12) 2 unspecified role unspecified role neutral component neutral component P15917:p.Asn173Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain N A Wed May 02 15:17:33 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asn173ala' Tue Jun 30 19:18:48 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asn173Ala' P15917:p.Lys176Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain K A Wed May 02 19:42:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys176ala' Tue Jun 30 16:25:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys176Ala' region Only the mutant proteins were radiolabeled at Met residues. 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P15917:p.Asp169Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D K Wed May 02 16:07:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp169lys' Tue Jun 30 19:52:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp169Lys' n-terminal LFN sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain P15917:p.Asp139Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D A Wed May 02 15:22:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp139ala' Tue Jun 30 16:21:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp139Ala' P15917:p.Tyr141Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain Y K Wed May 02 18:12:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr141lys' Tue Jun 30 17:11:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr141Lys' P15917:p.Glu168Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E A Wed May 02 19:55:37 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu168ala' Tue Jun 30 19:58:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu168Ala' P15917:p.Glu168Lys mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain E K Wed May 02 20:21:05 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu168lys' Tue Jun 30 15:45:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu168Lys' P15917:p.Asp169Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain D A Wed May 02 16:29:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'asp169ala' Tue Jun 30 18:00:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Asp169Ala' ecoli Escherichia coli (strain K12) physical association physical association When two mutated, oligomerization-deficient forms of PA are combined in the presence of LFN (31), a ternary complex is formed containing one molecule of each species. A paper describing the plasmid pET22b-PA indicates that PA is only 735 residues long but the Uniprot entry is 764 residues. This paper does not specify the length. The plasmid pET22b-PA does not contain the sequence coding for the signal peptide. Thus the mutations described in the paper (asp512lys and lys199glu&arg468ala&arg470asp) are mapped to residues 29 residues later in the Uniprot entry. They first examined LFN bound to the heptameric pre-pore, but because of the inherent tendency of these complexes to aggregate at high concentrations, they turned to a dimeric formof PA as an alternative binding partner for LFN. When two mutated,oligomerization-deficient forms of PA are combined in the presence of LFN, a ternary complex is formed containing one molecule of each species. PA was activated by incubation with trypsin at a final trypsin. Four discrete sites in LFN (i.e. residues 95�120, DX site 1; 137�147, DX site 2; 177�189, DX site 3; and 223�235, DX site 4) became significantly more protected from solvent exchange in the presence of PA. 2 and 3 Mapping all of the binding-defective mutants onto the four sites on the structure of LFN revealed the breadth of the binding surface defined by residues in DX sites 2 and 3. The distance between the most distal bindingdefective mutants, Asp-182 on DX site 3 to Glu-135 on DX site 2, was seen to be 40 A. The mutations in LFN listed above are those the authors specify as causing a defect in binding, however, from Fig. 3 A it seems that some of the other mutations did cause a decrease in binding.