ID PAG_BACAN Reviewed; 764 AA. AC P13423; Q937W2; Q937W3; Q9F5R7; Q9KH69; Q9RQU2; DT 01-JAN-1990, integrated into UniProtKB/Swiss-Prot. DT 18-OCT-2001, sequence version 2. DT 22-APR-2020, entry version 190. DE RecName: Full=Protective antigen; DE Short=PA; DE AltName: Full=Anthrax toxins translocating protein; DE AltName: Full=PA-83; DE Short=PA83; DE Contains: DE RecName: Full=Protective antigen PA-20; DE Short=PA20; DE Contains: DE RecName: Full=Protective antigen PA-63; DE Short=PA63; DE Flags: Precursor; GN Name=pagA; Synonyms=pag; GN OrderedLocusNames=pXO1-110, BXA0164, GBAA_pXO1_0164; OS Bacillus anthracis. OG Plasmid pXO1. OC Bacteria; Firmicutes; Bacilli; Bacillales; Bacillaceae; Bacillus; OC Bacillus cereus group. OX NCBI_TaxID=1392; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RX PubMed=3148491; DOI=10.1016/0378-1119(88)90439-8; RA Welkos S.L., Lowe J.R., Eden-Mccutchan F., Vodkin M., Leppla S.H., RA Schmidt J.J.; RT "Sequence and analysis of the DNA encoding protective antigen of Bacillus RT anthracis."; RL Gene 69:287-300(1988). RN [2] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=28, 33, BA1024, and BA1035; RX PubMed=10197996; RA Price L.B., Hugh-Jones M., Jackson P.J., Keim P.; RT "Genetic diversity in the protective antigen gene of Bacillus anthracis."; RL J. Bacteriol. 181:2358-2362(1999). RN [3] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=V770-NP1-R / ATCC 14185; RX PubMed=10899854; DOI=10.1128/iai.68.8.4549-4558.2000; RA Cohen S., Mendelson I., Altboum Z., Kobiler D., Elhanany E., Bino T., RA Leitner M., Inbar I., Rosenberg H., Gozes Y., Barak R., Fisher M., RA Kronman C., Velan B., Shafferman A.; RT "Attenuated nontoxinogenic and nonencapsulated recombinant Bacillus RT anthracis spore vaccines protect against anthrax."; RL Infect. Immun. 68:4549-4558(2000). RN [4] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=Sterne; RX PubMed=10515943; RA Okinaka R.T., Cloud K., Hampton O., Hoffmaster A.R., Hill K.K., Keim P., RA Koehler T.M., Lamke G., Kumano S., Mahillon J., Manter D., Martinez Y., RA Ricke D., Svensson R., Jackson P.J.; RT "Sequence and organization of pXO1, the large Bacillus anthracis plasmid RT harboring the anthrax toxin genes."; RL J. Bacteriol. 181:6509-6515(1999). RN [5] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=Ames / isolate Florida / A2012; RX PubMed=12004073; DOI=10.1126/science.1071837; RA Read T.D., Salzberg S.L., Pop M., Shumway M.F., Umayam L., Jiang L., RA Holtzapple E., Busch J.D., Smith K.L., Schupp J.M., Solomon D., Keim P., RA Fraser C.M.; RT "Comparative genome sequencing for discovery of novel polymorphisms in RT Bacillus anthracis."; RL Science 296:2028-2033(2002). RN [6] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=Ames ancestor; RX PubMed=18952800; DOI=10.1128/jb.01347-08; RA Ravel J., Jiang L., Stanley S.T., Wilson M.R., Decker R.S., Read T.D., RA Worsham P., Keim P.S., Salzberg S.L., Fraser-Liggett C.M., Rasko D.A.; RT "The complete genome sequence of Bacillus anthracis Ames 'Ancestor'."; RL J. Bacteriol. 191:445-446(2009). RN [7] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 9-751. RC STRAIN=Carbosap, and Ferrara; RX PubMed=12067380; DOI=10.1046/j.1365-2672.2002.01660.x; RA Adone R., Pasquali P., La Rosa G., Marianelli C., Muscillo M., RA Fasanella A., Francia M., Ciuchini F.; RT "Sequence analysis of the genes encoding for the major virulence factors of RT Bacillus anthracis vaccine strain 'Carbosap'."; RL J. Appl. Microbiol. 93:117-121(2002). RN [8] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 195-434. RC STRAIN=PAI; RX PubMed=14985634; RA Inoue S., Noguchi A., Tanabayashi K., Yamada A.; RT "Preparation of a positive control DNA for molecular diagnosis of Bacillus RT anthracis."; RL Jpn. J. Infect. Dis. 57:29-32(2004). RN [9] RP DOMAINS. RX PubMed=1651334; RA Singh Y., Klimpel K.R., Quinn C.P., Chaudhary V.K., Leppla S.H.; RT "The carboxyl-terminal end of protective antigen is required for receptor RT binding and anthrax toxin activity."; RL J. Biol. Chem. 266:15493-15497(1991). RN [10] RP CHARACTERIZATION. RC STRAIN=Sterne; RX PubMed=8051159; RA Milne J.C., Furlong D., Hanna P.C., Wall J.S., Collier R.J.; RT "Anthrax protective antigen forms oligomers during intoxication of RT mammalian cells."; RL J. Biol. Chem. 269:20607-20612(1994). RN [11] RP CHARACTERIZATION. RX PubMed=11207581; DOI=10.1046/j.1462-5822.2000.00052.x; RA Beauregard K.E., Collier R.J., Swanson J.A.; RT "Proteolytic activation of receptor-bound anthrax protective antigen on RT macrophages promotes its internalization."; RL Cell. Microbiol. 2:251-258(2000). RN [12] RP TOXIN REGULATION. RC STRAIN=Weybridge; RX PubMed=8300513; DOI=10.1128/jb.176.3.586-595.1994; RA Koehler T.M., Dai Z., Kaufman-Yarbray M.; RT "Regulation of the Bacillus anthracis protective antigen gene: CO2 and a RT trans-acting element activate transcription from one of two promoters."; RL J. Bacteriol. 176:586-595(1994). RN [13] RP FOLDING BY PSRA. RX PubMed=12606539; DOI=10.1074/jbc.m301244200; RA Williams R.C., Rees M.L., Jacobs M.F., Pragai Z., Thwaite J.E., RA Baillie L.W., Emmerson P.T., Harwood C.R.; RT "Production of Bacillus anthracis protective antigen is dependent on the RT extracellular chaperone, PrsA."; RL J. Biol. Chem. 278:18056-18062(2003). RN [14] RP INTERACTION WITH ANTHRAX TOXIN RECEPTOR. RX PubMed=14507921; DOI=10.1074/jbc.m307900200; RA Bradley K.A., Mogridge J., Jonah G., Rainey G.J.A., Batty S., Young J.A.T.; RT "Binding of anthrax toxin to its receptor is similar to alpha integrin- RT ligand interactions."; RL J. Biol. Chem. 278:49342-49347(2003). RN [15] RP MUTAGENESIS OF 342-PHE-PHE-343 AND ASP-344. RC STRAIN=Sterne; RX PubMed=7961869; RA Singh Y., Klimpel K.R., Arora N., Sharma M., Leppla S.H.; RT "The chymotrypsin-sensitive site, FFD315, in anthrax toxin protective RT antigen is required for translocation of lethal factor."; RL J. Biol. Chem. 269:29039-29046(1994). RN [16] RP MUTAGENESIS OF DOMAIN 4 LOOPS. RC STRAIN=Sterne; RX PubMed=10085028; RA Varughese M., Teixeira A.V., Liu S., Leppla S.H.; RT "Identification of a receptor-binding region within domain 4 of the RT protective antigen component of anthrax toxin."; RL Infect. Immun. 67:1860-1865(1999). RN [17] RP MUTAGENESIS OF TRP-375; MET-379 AND LEU-381. RC STRAIN=Sterne; RX PubMed=11178978; DOI=10.1006/bbrc.2001.4320; RA Batra S., Gupta P., Chauhan V., Singh A., Bhatnagar R.; RT "Trp 346 and Leu 352 residues in protective antigen are required for the RT expression of anthrax lethal toxin activity."; RL Biochem. Biophys. Res. Commun. 281:186-192(2001). RN [18] RP MUTAGENESIS OF PHE-581; PHE-583; ILE-591; LEU-595 AND ILE-603. RC STRAIN=Sterne; RX PubMed=11554763; DOI=10.1006/bbrc.2001.5613; RA Ahuja N., Kumar P., Bhatnagar R.; RT "Hydrophobic residues Phe552, Phe554, Ile562, Leu566, and Ile574 are RT required for oligomerization of anthrax protective antigen."; RL Biochem. Biophys. Res. Commun. 287:542-549(2001). RN [19] RP MUTAGENESIS OF PRO-289. RC STRAIN=Sterne; RX PubMed=11356563; DOI=10.1111/j.1574-6968.2001.tb10646.x; RA Khanna H., Chopra A.P., Arora N., Chaudhry A., Singh Y.; RT "Role of residues constituting the 2beta1 strand of domain II in the RT biological activity of anthrax protective antigen."; RL FEMS Microbiol. Lett. 199:27-31(2001). RN [20] RP MUTAGENESIS OF GLN-512; ASP-541; LEU-543 AND ARG-621. RX PubMed=11222612; DOI=10.1128/jb.183.6.2111-2116.2001; RA Mogridge J., Mourez M., Collier R.J.; RT "Involvement of domain 3 in oligomerization by the protective antigen RT moiety of anthrax toxin."; RL J. Bacteriol. 183:2111-2116(2001). RN [21] RP MUTAGENESIS OF LYS-426; ASP-454 AND PHE-456. RX PubMed=11113126; DOI=10.1074/jbc.m008309200; RA Sellman B.R., Nassi S., Collier R.J.; RT "Point mutations in anthrax protective antigen that block translocation."; RL J. Biol. Chem. 276:8371-8376(2001). RN [22] RP MUTAGENESIS OF PRO-213; LEU-216; PHE-231; LEU-232; PRO-234; ILE-236; RP ILE-239; TRP-255 AND PHE-265. RC STRAIN=Sterne; RX PubMed=12117959; DOI=10.1128/iai.70.8.4477-4484.2002; RA Chauhan V., Bhatnagar R.; RT "Identification of amino acid residues of anthrax protective antigen RT involved in binding with lethal factor."; RL Infect. Immun. 70:4477-4484(2002). RN [23] RP MUTAGENESIS OF ILE-393; THR-409; SER-411; THR-422; LYS-426; ASN-428; RP TYR-440; ASN-451; ASP-454 AND PHE-456. RX PubMed=14623961; DOI=10.1073/pnas.2436299100; RA Mourez M., Yan M., Lacy D.B., Dillon L., Bentsen L., Marpoe A., Maurin C., RA Hotze E., Wigelsworth D., Pimental R.-A., Ballard J.D., Collier R.J., RA Tweten R.K.; RT "Mapping dominant-negative mutations of anthrax protective antigen by RT scanning mutagenesis."; RL Proc. Natl. Acad. Sci. U.S.A. 100:13803-13808(2003). RN [24] RP MUTAGENESIS OF ASN-686; LYS-708; LYS-709; TYR-710; ASN-711; ASP-712; RP LYS-713; LEU-714; PRO-715; LEU-716; TYR-717; ILE-718; ASN-720; PRO-721 AND RP ASN-722. RX PubMed=12771151; DOI=10.1074/jbc.m301154200; RA Rosovitz M.J., Schuck P., Varughese M., Chopra A.P., Mehra V., Singh Y., RA McGinnis L.M., Leppla S.H.; RT "Alanine-scanning mutations in domain 4 of anthrax toxin protective antigen RT reveal residues important for binding to the cellular receptor and to a RT neutralizing monoclonal antibody."; RL J. Biol. Chem. 278:30936-30944(2003). RN [25] RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS). RX PubMed=9039918; DOI=10.1038/385833a0; RA Petosa C., Collier R.J., Klimpel K.R., Leppla S.H., Liddington R.C.; RT "Crystal structure of the anthrax toxin protective antigen."; RL Nature 385:833-838(1997). RN [26] RP X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) OF 30-764 IN COMPLEX WITH ANTXR2. RX PubMed=15243628; DOI=10.1038/nature02763; RA Santelli E., Bankston L.A., Leppla S.H., Liddington R.C.; RT "Crystal structure of a complex between anthrax toxin and its host cell RT receptor."; RL Nature 430:905-908(2004). RN [27] RP X-RAY CRYSTALLOGRAPHY (4.3 ANGSTROMS) OF 203-764 IN COMPLEX WITH ANTXR2. RX PubMed=15326297; DOI=10.1073/pnas.0405405101; RA Lacy D.B., Wigelsworth D.J., Melnyk R.A., Harrison S.C., Collier R.J.; RT "Structure of heptameric protective antigen bound to an anthrax toxin RT receptor: a role for receptor in pH-dependent pore formation."; RL Proc. Natl. Acad. Sci. U.S.A. 101:13147-13151(2004). RN [28] RP REVIEW. RX PubMed=11544370; DOI=10.1146/annurev.micro.55.1.647; RA Mock M., Fouet A.; RT "Anthrax."; RL Annu. Rev. Microbiol. 55:647-671(2001). CC -!- FUNCTION: One of the three proteins composing the anthrax toxin, the CC agent which infects many mammalian species and that may cause death. PA CC binds to a receptor (ATR) in sensitive eukaryotic cells, thereby CC facilitating the translocation of the enzymatic toxin components, edema CC factor and lethal factor, across the target cell membrane. PA CC associated with LF causes death when injected, PA associated with EF CC produces edema. PA induces immunity to infection with anthrax. CC -!- SUBUNIT: Anthrax toxins are composed of three distinct proteins, a CC protective antigen (PA), a lethal factor (LF) and an edema factor (EF). CC None of these is toxic by itself. PA+LF forms the lethal toxin (LeTx); CC PA+EF forms the edema toxin (EdTx). PA-63 forms heptamers and this CC oligomerization is required for LF or EF binding. This complex is CC endocytosed by the host. Once activated, at low pH, the heptamer CC undergoes conformational changes and converts from prepore to pore CC inserted in the membrane, forming cation-selective channels and CC triggering the release of LF and EF in the host cytoplasm. CC {ECO:0000269|PubMed:15243628, ECO:0000269|PubMed:15326297}. CC -!- INTERACTION: CC P13423; P13423; NbExp=16; IntAct=EBI-456868, EBI-456868; CC P13423; Q9H6X2-2: ANTXR1; Xeno; NbExp=3; IntAct=EBI-456868, EBI-905659; CC P13423; P58335: ANTXR2; Xeno; NbExp=7; IntAct=EBI-456868, EBI-456840; CC P13423; P0A6F5: groL; Xeno; NbExp=2; IntAct=EBI-456868, EBI-543750; CC P13423; P15917: lef; NbExp=28; IntAct=EBI-456868, EBI-456923; CC -!- SUBCELLULAR LOCATION: Secreted. Note=Secreted through the Sec-dependent CC secretion pathway. Therefore, PA is translocated across the membrane in CC an unfolded state and then it is folded into its native configuration CC on the trans side of the membrane, prior to its release to the CC environment. PA requires the extracellular chaperone PrsA for efficient CC folding. {ECO:0000269|PubMed:12606539}. CC -!- DOMAIN: The molecule is folded into four functional domains. Each CC domain is required for a particular step in the toxicity process. CC Domain 1 contains two calcium ions and the proteolytic activation site. CC Cleavage of the PA monomer releases the subdomain 1a, which is the N- CC terminal fragment of 20-kDa (PA20). The subdomain 1b is part of the CC remaining 63-kDa fragment (PA63) and contains the binding sites for LP CC and EF. Domain 2 is a beta-barrel core containing a large flexible loop CC that has been implicated in membrane insertion and pore formation. CC There is a chymotrypsin cleavage site in this loop that is required for CC toxicity. Domain 3 has a hydrophobic patch thought to be involved in CC protein-protein interactions. Domain 4 appears to be a separate domain CC and shows limited contact with the other three domains: it would swing CC out of the way during membrane insertion. It is required for binding to CC the receptor; the small loop is involved in receptor recognition. CC {ECO:0000269|PubMed:1651334}. CC -!- PTM: Proteolytic activation by furin or a furin-like protease cleaves CC the protein in two parts, PA-20 and PA-63; the latter is the mature CC protein. The cleavage occurs at the cell surface and probably in the CC serum of infected animals as well; both native and cleaved PA are able CC to bind to the cell receptor. The release of PA20 from the remaining CC receptor-bound PA63 exposes the binding site for EF and LF, and CC promotes oligomerization and internalization of the protein. CC -!- MISCELLANEOUS: In PubMed:10085028 multiple mutagenesis experiments were CC performed that showed that the residues present in the small loop of CC domain 4, and not the ones in the large loop, are involved in receptor CC recognition. In PubMed:14623961 high-throughput scanning mutagenesis CC experiments were performed in which all residues of PA-63 were mutated CC into Cys. Dominantly negative (DN) mutants were all clustered in domain CC 2. DN mutants prevent the conformational transition of PA-63 from the CC prepore to the pore state. CC -!- SIMILARITY: Belongs to the bacterial binary toxin family. CC {ECO:0000305}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; M22589; AAA22637.1; -; Genomic_DNA. DR EMBL; AF306778; AAG24446.1; -; Genomic_DNA. DR EMBL; AF306779; AAG24447.1; -; Genomic_DNA. DR EMBL; AF306780; AAG24448.1; -; Genomic_DNA. DR EMBL; AF306781; AAG24449.1; -; Genomic_DNA. DR EMBL; AF306782; AAG24450.1; -; Genomic_DNA. DR EMBL; AF306783; AAG24451.1; -; Genomic_DNA. DR EMBL; AF268967; AAF86457.1; -; Genomic_DNA. DR EMBL; AF065404; AAD32414.1; -; Genomic_DNA. DR EMBL; AE011190; AAM26109.1; -; Genomic_DNA. DR EMBL; AE017336; AAT28905.2; -; Genomic_DNA. DR EMBL; AJ413936; CAC93934.1; -; Genomic_DNA. DR EMBL; AJ413937; CAC93935.1; -; Genomic_DNA. DR EMBL; AB125961; BAD14937.1; -; Genomic_DNA. DR PIR; I39934; I39934. DR RefSeq; NP_052806.1; NC_001496.1. DR RefSeq; WP_000746486.1; NZ_QPKQ01000017.1. DR RefSeq; WP_000746487.1; NZ_QAEM01000007.1. DR RefSeq; WP_000746488.1; NZ_QANP01000007.1. DR PDB; 1ACC; X-ray; 2.10 A; A=30-764. DR PDB; 1T6B; X-ray; 2.50 A; X=30-764. DR PDB; 1TX5; Model; -; C=30-764. DR PDB; 1TZN; X-ray; 4.30 A; A/B/C/D/E/F/G/H/I/J/K/L/M/O=203-764. DR PDB; 1TZO; X-ray; 3.60 A; A/B/C/D/E/F/G/H/I/J/K/L/M/O=203-764. DR PDB; 1V36; Model; -; A/B/C/D/E/F/G=197-764. DR PDB; 3ETB; X-ray; 3.80 A; J/K/L/M=621-764. DR PDB; 3INO; X-ray; 1.95 A; A/B=624-764. DR PDB; 3J9C; EM; 2.90 A; A=203-764. DR PDB; 3KWV; X-ray; 3.10 A; A/B/D/E=197-764. DR PDB; 3MHZ; X-ray; 1.70 A; A=30-764. DR PDB; 3Q8A; X-ray; 3.13 A; A=30-764. DR PDB; 3Q8B; X-ray; 2.00 A; A=30-764. DR PDB; 3Q8C; X-ray; 2.85 A; A=30-764. DR PDB; 3Q8E; X-ray; 2.10 A; A=30-764. DR PDB; 3Q8F; X-ray; 2.10 A; A=30-764. DR PDB; 3TEW; X-ray; 1.45 A; A=30-764. DR PDB; 3TEX; X-ray; 1.70 A; A=30-764. DR PDB; 3TEY; X-ray; 2.12 A; A=30-764. DR PDB; 3TEZ; X-ray; 1.83 A; A=30-764. DR PDB; 4EE2; X-ray; 1.91 A; A=30-764. DR PDB; 4H2A; X-ray; 1.62 A; A=30-764. DR PDB; 4NAM; X-ray; 1.70 A; A=30-764. DR PDB; 5FR3; X-ray; 1.94 A; A=43-764. DR PDBsum; 1ACC; -. DR PDBsum; 1T6B; -. DR PDBsum; 1TX5; -. DR PDBsum; 1TZN; -. DR PDBsum; 1TZO; -. DR PDBsum; 1V36; -. DR PDBsum; 3ETB; -. DR PDBsum; 3INO; -. DR PDBsum; 3J9C; -. DR PDBsum; 3KWV; -. DR PDBsum; 3MHZ; -. DR PDBsum; 3Q8A; -. DR PDBsum; 3Q8B; -. DR PDBsum; 3Q8C; -. DR PDBsum; 3Q8E; -. DR PDBsum; 3Q8F; -. DR PDBsum; 3TEW; -. DR PDBsum; 3TEX; -. DR PDBsum; 3TEY; -. DR PDBsum; 3TEZ; -. DR PDBsum; 4EE2; -. DR PDBsum; 4H2A; -. DR PDBsum; 4NAM; -. DR PDBsum; 5FR3; -. DR SMR; P13423; -. DR DIP; DIP-29841N; -. DR IntAct; P13423; 16. DR MINT; P13423; -. DR BindingDB; P13423; -. DR ChEMBL; CHEMBL5352; -. DR DrugBank; DB09057; Anthrax immune globulin human. DR DrugBank; DB08902; Raxibacumab. DR DrugCentral; P13423; -. DR TCDB; 1.C.42.1.1; the channel-forming bacillus anthracis protective antigen (bapa) family. DR PRIDE; P13423; -. DR ABCD; P13423; -. DR EnsemblBacteria; AAM26109; AAM26109; BX_A0164. DR EnsemblBacteria; AAT28905; AAT28905; GBAA_pXO1_0164. DR GeneID; 3361714; -. DR GeneID; 39691010; -. DR KEGG; bar:GBAA_pXO1_0164; -. DR HOGENOM; CLU_015269_0_0_9; -. DR KO; K11030; -. DR BioCyc; ANTHRA:GBAA_PXO1_0164-MONOMER; -. DR Reactome; R-HSA-5210891; Uptake and function of anthrax toxins. DR EvolutionaryTrace; P13423; -. DR PHI-base; PHI:4090; -. DR PRO; PR:P13423; -. DR Proteomes; UP000000594; Plasmid pXO1. DR GO; GO:0005576; C:extracellular region; IEA:UniProtKB-SubCell. DR GO; GO:0042802; F:identical protein binding; IPI:IntAct. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0090729; F:toxin activity; IEA:UniProtKB-KW. DR GO; GO:0010629; P:negative regulation of gene expression; IGI:UniProtKB. DR GO; GO:0043409; P:negative regulation of MAPK cascade; IGI:UniProtKB. DR GO; GO:0001933; P:negative regulation of protein phosphorylation; IGI:UniProtKB. DR GO; GO:0000122; P:negative regulation of transcription by RNA polymerase II; IGI:UniProtKB. DR GO; GO:0009405; P:pathogenesis; IEA:UniProtKB-KW. DR GO; GO:0044533; P:positive regulation of apoptotic process in other organism; IGI:UniProtKB. DR GO; GO:0051260; P:protein homooligomerization; IEA:InterPro. DR GO; GO:0035897; P:proteolysis in other organism; IGI:UniProtKB. DR GO; GO:1903140; P:regulation of establishment of endothelial barrier; IGI:UniProtKB. DR GO; GO:0061136; P:regulation of proteasomal protein catabolic process; IGI:UniProtKB. DR Gene3D; 2.60.120.240; -; 1. DR InterPro; IPR003896; Bacterial_exotoxin_B. DR InterPro; IPR035331; Binary_toxB_3. DR InterPro; IPR037524; PA14/GLEYA. DR InterPro; IPR011658; PA14_dom. DR InterPro; IPR035088; PA_Ca-bd. DR InterPro; IPR027439; PA_heptamer_dom. DR InterPro; IPR037149; PA_heptamer_dom_sf. DR Pfam; PF03495; Binary_toxB; 1. DR Pfam; PF17475; Binary_toxB_2; 1. DR Pfam; PF17476; Binary_toxB_3; 1. DR Pfam; PF07691; PA14; 1. DR PRINTS; PR01391; BINARYTOXINB. DR SMART; SM00758; PA14; 1. DR PROSITE; PS51820; PA14; 1. PE 1: Evidence at protein level; KW 3D-structure; Calcium; Cleavage on pair of basic residues; Metal-binding; KW Plasmid; Reference proteome; Secreted; Signal; Toxin; Virulence. FT SIGNAL 1..29 FT CHAIN 30..764 FT /note="Protective antigen" FT /id="PRO_0000021996" FT CHAIN 30..196 FT /note="Protective antigen PA-20" FT /id="PRO_0000021997" FT CHAIN 197..764 FT /note="Protective antigen PA-63" FT /id="PRO_0000021998" FT DOMAIN 43..179 FT /note="PA14" FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01164" FT REGION 30..287 FT /note="Domain 1, calcium-binding; LF and EF binding sites" FT REGION 288..516 FT /note="Domain 2, membrane insertion and heptamerization" FT REGION 517..624 FT /note="Domain 3, heptamerization" FT REGION 625..764 FT /note="Domain 4, binding to the receptor" FT METAL 206 FT /note="Calcium" FT METAL 208 FT /note="Calcium" FT METAL 210 FT /note="Calcium" FT METAL 217 FT /note="Calcium" FT SITE 196..197 FT /note="Cleavage; by furin" FT SITE 343..344 FT /note="Cleavage; by chymotrypsin; required for FT translocation of LF and EF" FT SITE 712 FT /note="Essential for binding to cell receptor" FT VARIANT 295 FT /note="M -> I (in strain: PAI)" FT VARIANT 392 FT /note="N -> D (in strain: PAI)" FT VARIANT 560 FT /note="F -> L (in Sverdlovsk sample)" FT VARIANT 565 FT /note="P -> S (in strain: BA1024)" FT VARIANT 600 FT /note="A -> V (in strain: BA1024, V770-NP1-R, Carbosap and FT Ferrara)" FT MUTAGEN 213 FT /note="P->A: Decrease in the ability to bind to LF and FT partially toxic at high concentrations." FT /evidence="ECO:0000269|PubMed:12117959" FT MUTAGEN 216 FT /note="L->A: Decrease in the ability to bind to LF and FT partially toxic at high concentrations." FT /evidence="ECO:0000269|PubMed:12117959" FT MUTAGEN 231 FT /note="F->A: Loss of ability to bind to LF and completely FT non-toxic." FT /evidence="ECO:0000269|PubMed:12117959" FT MUTAGEN 232 FT /note="L->A: Loss of ability to bind to LF and completely FT non-toxic." FT /evidence="ECO:0000269|PubMed:12117959" FT MUTAGEN 234 FT /note="P->A: Loss of ability to bind to LF and completely FT non-toxic." FT /evidence="ECO:0000269|PubMed:12117959" FT MUTAGEN 236 FT /note="I->A: Loss of ability to bind to LF and completely FT non-toxic." FT /evidence="ECO:0000269|PubMed:12117959" FT MUTAGEN 239 FT /note="I->A: Decrease in the ability to bind to LF and FT partially toxic at high concentrations." FT /evidence="ECO:0000269|PubMed:12117959" FT MUTAGEN 255 FT /note="W->A: No effect on LF-binding ability and as toxic FT as the wild-type." FT /evidence="ECO:0000269|PubMed:12117959" FT MUTAGEN 265 FT /note="F->A: No effect on LF-binding ability and as toxic FT as the wild-type." FT /evidence="ECO:0000269|PubMed:12117959" FT MUTAGEN 289 FT /note="P->A: Reduced toxicity in combination with lethal FT factor. Decreased membrane insertion and translocation of FT LF." FT /evidence="ECO:0000269|PubMed:11356563" FT MUTAGEN 342..344 FT /note="FFD->AAA: Decrease in toxicity probably due to slow FT translocation of LF." FT /evidence="ECO:0000269|PubMed:10085028" FT MUTAGEN 342..343 FT /note="Missing: Loss of toxicity probably due to loss of FT capability to translocate LF." FT /evidence="ECO:0000269|PubMed:7961869" FT MUTAGEN 342 FT /note="F->C: Loss of toxicity probably due to loss of FT capability to translocate LF." FT /evidence="ECO:0000269|PubMed:10085028" FT MUTAGEN 344 FT /note="D->A: Decrease in toxicity probably due to slow FT translocation of LF." FT /evidence="ECO:0000269|PubMed:7961869" FT MUTAGEN 375 FT /note="W->A: Loss of toxicity probably due to faulty FT membrane insertion or translocation of LF/EF into the FT cytosol." FT /evidence="ECO:0000269|PubMed:11178978" FT MUTAGEN 379 FT /note="M->A: No effect." FT /evidence="ECO:0000269|PubMed:11178978" FT MUTAGEN 381 FT /note="L->A: Loss of toxicity probably due to faulty FT membrane insertion or translocation of LF/EF into the FT cytosol." FT /evidence="ECO:0000269|PubMed:11178978" FT MUTAGEN 393 FT /note="I->C: Loss of capability to undergo conformational FT changes that lead to pore formation and translocation." FT /evidence="ECO:0000269|PubMed:14623961" FT MUTAGEN 409 FT /note="T->C: Loss of capability to undergo conformational FT changes that lead to pore formation and translocation." FT /evidence="ECO:0000269|PubMed:14623961" FT MUTAGEN 411 FT /note="S->C: Loss of capability to undergo conformational FT changes that lead to pore formation and translocation." FT /evidence="ECO:0000269|PubMed:14623961" FT MUTAGEN 422 FT /note="T->C: Loss of capability to undergo conformational FT changes that lead to pore formation and translocation." FT /evidence="ECO:0000269|PubMed:14623961" FT MUTAGEN 426 FT /note="K->A,D: Loss of capability to undergo conformational FT changes that lead to pore formation and translocation." FT /evidence="ECO:0000269|PubMed:11113126, FT ECO:0000269|PubMed:14623961" FT MUTAGEN 428 FT /note="N->C: Loss of capability to undergo conformational FT changes that lead to pore formation and translocation." FT /evidence="ECO:0000269|PubMed:14623961" FT MUTAGEN 440 FT /note="Y->C: Loss of capability to undergo conformational FT changes that lead to pore formation and translocation." FT /evidence="ECO:0000269|PubMed:14623961" FT MUTAGEN 451 FT /note="N->C: Loss of capability to undergo conformational FT changes that lead to pore formation and translocation." FT /evidence="ECO:0000269|PubMed:14623961" FT MUTAGEN 454 FT /note="D->A,K: Loss of capability to undergo conformational FT changes that lead to pore formation and translocation." FT /evidence="ECO:0000269|PubMed:11113126, FT ECO:0000269|PubMed:14623961" FT MUTAGEN 456 FT /note="F->A: Loss of capability to undergo conformational FT changes that lead to pore formation and translocation." FT /evidence="ECO:0000269|PubMed:11113126, FT ECO:0000269|PubMed:14623961" FT MUTAGEN 512 FT /note="Q->A: Loss of heptamerization capability." FT /evidence="ECO:0000269|PubMed:11222612" FT MUTAGEN 541 FT /note="D->A: Loss of heptamerization capability." FT /evidence="ECO:0000269|PubMed:11222612" FT MUTAGEN 543 FT /note="L->A: Decrease in heptamerization capability." FT /evidence="ECO:0000269|PubMed:11222612" FT MUTAGEN 581 FT /note="F->A: Loss of toxicity due to defective FT oligomerization." FT /evidence="ECO:0000269|PubMed:11554763" FT MUTAGEN 583 FT /note="F->A: Decrease in toxicity due to defective FT oligomerization." FT /evidence="ECO:0000269|PubMed:11554763" FT MUTAGEN 591 FT /note="I->A: Loss of toxicity due to defective FT oligomerization." FT /evidence="ECO:0000269|PubMed:11554763" FT MUTAGEN 595 FT /note="L->A: Loss of toxicity due to defective FT oligomerization." FT /evidence="ECO:0000269|PubMed:11554763" FT MUTAGEN 603 FT /note="I->A: Loss of toxicity due to defective FT oligomerization." FT /evidence="ECO:0000269|PubMed:11554763" FT MUTAGEN 621 FT /note="R->A: No effect." FT /evidence="ECO:0000269|PubMed:11222612" FT MUTAGEN 686 FT /note="N->A: Decrease in toxicity due to decrease in cell FT binding." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 708 FT /note="K->A: No effect on toxicity." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 709 FT /note="K->A: Slight decrease in toxicity." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 710 FT /note="Y->A: Great decrease in toxicity due to decrease in FT cell binding." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 711 FT /note="N->A: Loss of toxicity due to decrease in cell FT binding." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 712 FT /note="D->A: Loss of toxicity due to decrease in cell FT binding." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 713 FT /note="K->A: No effect on toxicity." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 714 FT /note="L->A: No effect on toxicity." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 715 FT /note="P->A: Great decrease in toxicity due to decrease in FT cell binding." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 716 FT /note="L->A: Decrease in toxicity due to decrease in cell FT binding." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 717 FT /note="Y->A: No effect on toxicity." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 718 FT /note="I->A: Decrease in toxicity due to decrease in cell FT binding." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 719 FT /note="S->A: No effect on toxicity." FT /evidence="ECO:0000269|PubMed:10085028" FT MUTAGEN 720 FT /note="N->A: No effect on toxicity." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 721 FT /note="P->A: No effect on toxicity." FT /evidence="ECO:0000269|PubMed:12771151" FT MUTAGEN 722 FT /note="N->A: No effect on toxicity." FT /evidence="ECO:0000269|PubMed:12771151" FT CONFLICT 314 FT /note="Q -> E (in Ref. 1; AAA22637)" FT /evidence="ECO:0000305" FT HELIX 41..44 FT /evidence="ECO:0000244|PDB:4H2A" FT STRAND 47..55 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 60..71 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 76..78 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 84..86 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 91..99 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 104..110 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 113..115 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 116..120 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 123..129 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 135..137 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 142..150 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 155..159 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 162..166 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 168..170 FT /evidence="ECO:0000244|PDB:1ACC" FT STRAND 172..174 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 177..179 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 186..188 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 191..193 FT /evidence="ECO:0000244|PDB:4H2A" FT STRAND 199..201 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 210..212 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 214..219 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 221..225 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 230..234 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 237..240 FT /evidence="ECO:0000244|PDB:3TEW" FT TURN 241..244 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 255..258 FT /evidence="ECO:0000244|PDB:3Q8A" FT STRAND 259..262 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 264..269 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 278..281 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 291..302 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 318..326 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 331..340 FT /evidence="ECO:0000244|PDB:3J9C" FT STRAND 345..354 FT /evidence="ECO:0000244|PDB:3J9C" FT STRAND 357..363 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 369..371 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 375..379 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 383..385 FT /evidence="ECO:0000244|PDB:3J9C" FT STRAND 386..397 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 399..401 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 403..405 FT /evidence="ECO:0000244|PDB:4EE2" FT STRAND 410..414 FT /evidence="ECO:0000244|PDB:3TEW" FT TURN 415..417 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 418..423 FT /evidence="ECO:0000244|PDB:3TEW" FT TURN 427..429 FT /evidence="ECO:0000244|PDB:3J9C" FT STRAND 438..441 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 448..451 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 456..458 FT /evidence="ECO:0000244|PDB:3KWV" FT STRAND 461..464 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 465..474 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 476..481 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 487..492 FT /evidence="ECO:0000244|PDB:3TEW" FT TURN 493..496 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 497..505 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 506..508 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 510..516 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 517..522 FT /evidence="ECO:0000244|PDB:3TEW" FT TURN 524..527 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 530..535 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 542..546 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 552..560 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 565..568 FT /evidence="ECO:0000244|PDB:3MHZ" FT HELIX 576..578 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 579..583 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 585..597 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 603..605 FT /evidence="ECO:0000244|PDB:3TEW" FT TURN 607..609 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 611..613 FT /evidence="ECO:0000244|PDB:3MHZ" FT STRAND 617..622 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 625..627 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 633..636 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 638..644 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 648..652 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 655..658 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 662..666 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 668..676 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 678..680 FT /evidence="ECO:0000244|PDB:3KWV" FT STRAND 682..684 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 689..691 FT /evidence="ECO:0000244|PDB:4EE2" FT STRAND 695..697 FT /evidence="ECO:0000244|PDB:3TEW" FT TURN 699..701 FT /evidence="ECO:0000244|PDB:3KWV" FT STRAND 703..708 FT /evidence="ECO:0000244|PDB:3TEW" FT TURN 709..713 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 723..731 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 732..734 FT /evidence="ECO:0000244|PDB:3TEW" FT STRAND 741..743 FT /evidence="ECO:0000244|PDB:1ACC" FT STRAND 753..759 FT /evidence="ECO:0000244|PDB:3TEW" FT HELIX 760..763 FT /evidence="ECO:0000244|PDB:3TEW" SQ SEQUENCE 764 AA; 85811 MW; 3AE1EFBF48FAA03F CRC64; MKKRKVLIPL MALSTILVSS TGNLEVIQAE VKQENRLLNE SESSSQGLLG YYFSDLNFQA PMVVTSSTTG DLSIPSSELE NIPSENQYFQ SAIWSGFIKV KKSDEYTFAT SADNHVTMWV DDQEVINKAS NSNKIRLEKG RLYQIKIQYQ RENPTEKGLD FKLYWTDSQN KKEVISSDNL QLPELKQKSS NSRKKRSTSA GPTVPDRDND GIPDSLEVEG YTVDVKNKRT FLSPWISNIH EKKGLTKYKS SPEKWSTASD PYSDFEKVTG RIDKNVSPEA RHPLVAAYPI VHVDMENIIL SKNEDQSTQN TDSQTRTISK NTSTSRTHTS EVHGNAEVHA SFFDIGGSVS AGFSNSNSST VAIDHSLSLA GERTWAETMG LNTADTARLN ANIRYVNTGT APIYNVLPTT SLVLGKNQTL ATIKAKENQL SQILAPNNYY PSKNLAPIAL NAQDDFSSTP ITMNYNQFLE LEKTKQLRLD TDQVYGNIAT YNFENGRVRV DTGSNWSEVL PQIQETTARI IFNGKDLNLV ERRIAAVNPS DPLETTKPDM TLKEALKIAF GFNEPNGNLQ YQGKDITEFD FNFDQQTSQN IKNQLAELNA TNIYTVLDKI KLNAKMNILI RDKRFHYDRN NIAVGADESV VKEAHREVIN SSTEGLLLNI DKDIRKILSG YIVEIEDTEG LKEVINDRYD MLNISSLRQD GKTFIDFKKY NDKLPLYISN PNYKVNVYAV TKENTIINPS ENGDTSTNGI KKILIFSKKG YEIG //