ID PAG_BACAN STANDARD; PRT; 764 AA. AC P13423; Q9RQU2; Q9F5R7; Q9KH69; DT 01-JAN-1990 (Rel. 13, Created) DT 16-OCT-2001 (Rel. 40, Last sequence update) DT 01-MAR-2002 (Rel. 41, Last annotation update) DE Protective antigen precursor (PA) (PA-83) (PA83) (Anthrax toxins DE translocating protein) [Contains: PA-20 (PA20); PA-63 (PA63)]. GN PAGA OR PAG OR PXO1-110. OS Bacillus anthracis. OG Plasmid pXO1. OC Bacteria; Firmicutes; Bacillus/Clostridium group; OC Bacillus/Staphylococcus group; Bacillus. OX NCBI_TaxID=1392; RN [1] RP SEQUENCE FROM N.A. RX MEDLINE=89172073; PubMed=3148491; 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 RT Bacillus anthracis."; RL Gene 69:287-300(1988). RN [2] RP SEQUENCE FROM N.A. RC STRAIN=28, 33, BA1024, AND BA1035; RX MEDLINE=99214082; PubMed=10197996; RA Price L.B., Hugh-Jones M., Jackson P.J., Keim P.; RT "Genetic diversity in the protective antigen gene of Bacillus RT anthracis."; RL J. Bacteriol. 181:2358-2362(1999). RN [3] RP SEQUENCE FROM N.A. RC STRAIN=V770-NP1-R / ATCC 14185; RX MEDLINE=20359347; PubMed=10899854; 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 SEQUENCE FROM N.A. RC STRAIN=Sterne; RX MEDLINE=99445483; PubMed=10515943; RA Okinaka R.T., Cloud K., Hampton O., Hoffmaster A.R., Hill K.K., RA Keim P., Koehler T.M., Lamke G., Kumano S., Mahillon J., Manter D., RA Martinez Y., Ricke D., Svensson R., Jackson P.J.; RT "Sequence and organization of pXO1, the large Bacillus anthracis RT plasmid harboring the Anthrax toxin genes."; RL J. Bacteriol. 181:6509-6515(1999). RN [5] RP DOMAINS. RX MEDLINE=91332080; 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 RT receptor binding and anthrax toxin activity."; RL J. Biol. Chem. 266:15493-15497(1991). RN [6] 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 [7] RP CHARACTERIZATION. RX PubMed=11207581; RA Beauregard K.E., Collier R.J., Swanson J.A.; RT "Proteolytic activation of receptor-bound anthrax protective antigen RT on macrophages promotes its internalization."; RL Cell. Microbiol. 2:251-258(2000). RN [8] RP TOXIN REGULATION. RC STRAIN=Weybridge; RX PubMed=8300513; RA Koehler T.M., Dai Z., Kaufman-Yarbray M.; RT "Regulation of the Bacillus anthracis protective antigen gene: CO2 and RT a trans-acting element activate transcription from one of two RT promoters."; RL J. Bacteriol. 176:586-595(1994). RN [9] RP MUTAGENESIS OF PHE-342; 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 [10] 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 [11] RP MUTAGENESIS OF TRP-375; MET-379 AND LEU-381. RC STRAIN=Sterne; RX PubMed=11178978; RA Batra S., Gupta P., Chauhan V., Singh A., Bhatnagar R.; RT "Trp 346 and Leu 352 residues in protective antigen are required for RT the expression of anthrax lethal toxin activity."; RL Biochem. Biophys. Res. Commun. 281:186-192(2001). RN [12] RP MUTAGENESIS OF PHE-581; PHE-583; ILE-591; LEU-595 AND ILE-603. RC STRAIN=Sterne; RX PubMed=11554763; 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 [13] RP MUTAGENESIS OF PRO-289. RC STRAIN=Sterne; RX PubMed=11356563; 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 [14] RP MUTAGENESIS OF GLN-512; ASP-541; LEU-543 AND ARG-621. RX PubMed=11222612; 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 [15] RP MUTAGENESIS OF LYS-426; ASP-454 AND PHE-456. RX PubMed=11113126; RA Sellman B.R., Nassi S., Collier R.J.; RT "Point mutations in anthrax protective antigen that block RT translocation."; RL J. Biol. Chem. 276:8371-8376(2001). RN [16] RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS). RX MEDLINE=97192099; PubMed=9039918; 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 [17] RP REVIEW. RX PubMed=11544370; 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, CC the agent which infects many mammalian species and that may cause CC death. PA binds to a receptor (ATR) in sensitive eukaryotic CC cells, thereby facilitating the translocation of the enzymatic CC toxin components, edema factor and lethal factor, across the CC target cell membrane. PA associated with LF causes death when CC injected, PA associated with EF produces edema. PA induces CC 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 CC (EF). None of these is toxic by itself. PA+LF forms the lethal CC toxin (LeTx); PA+EF forms the edema toxin (EdTx). PA-63 forms CC heptamers and this oligomerization is required for LF or EF CC binding. Once activated, at low pH, the heptamer undergoes CC conformational changes and converts from prepore to pore inserted CC in the membrane, forming cation-selective channels. CC -!- SUBCELLULAR LOCATION: Secreted. 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 CC site. Cleavage of the PA monomer releases the subdomain 1a, which CC is the N-terminal fragment of 20-kDa (PA20). The subdomain 1b is CC part of the remaining 63-kDa fragment (PA63). Domain 2 is a beta- CC barrel core containing a large flexible loop that has been CC implicated in membrane insertion and pore formation. There is a CC chymotrypsin cleavage site in this loop that is required for CC toxicity. Domain 3 has a hydrophobic patch thought to be involved CC in protein-protein interactions. Domain 4 appears to be a separate CC domain and shows limited contact with the other three domains: it CC would swing out of the way during membrane insertion. Domain 4 CC contains the binding sites for LP and EF and it is required for CC binding to the receptor; the small loop is involved in receptor CC recognition. CC -!- PTM: Proteolytic activation by furin or a furin-like protease CC cleaves the protein in two parts, PA-20 and PA-63; the latter is CC the mature protein. The cleavage occurs at the cell surface and CC probably in the serum of infected animals as well; both native and CC cleaved PA are able to bind to the cell receptor. The release of CC PA20 from the remaining receptor-bound PA63 exposes the binding CC site for EF and LF, and promotes oligomerization and CC internalization of the protein. CC -!- MISCELLANEOUS: In Ref.9 multiple mutagenesis experiments were CC performed that showed that the residues present in the small loop CC of domain 4, and not the ones in the large loop, are involved in CC receptor recognition. CC -!- SIMILARITY: BELONGS TO THE BACTERIAL BINARY TOXIN FAMILY. 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; -. DR EMBL; AF306778; AAG24446.1; -. DR EMBL; AF306779; AAG24447.1; -. DR EMBL; AF306780; AAG24448.1; -. DR EMBL; AF306781; AAG24449.1; -. DR EMBL; AF306782; AAG24450.1; -. DR EMBL; AF306783; AAG24451.1; -. DR EMBL; AF268967; AAF86457.1; -. DR EMBL; AF065404; AAD32414.1; -. DR PDB; 1ACC; 11-FEB-98. DR InterPro; IPR003896; Binary_toxB. DR PRINTS; PR01391; BINARYTOXINB. KW Toxin; Virulence; Calcium-binding; Signal; Plasmid; 3D-structure. FT SIGNAL 1 29 FT CHAIN 30 764 PROTECTIVE ANTIGEN. FT CHAIN 30 196 PROTECTIVE ANTIGEN, PA-20. FT CHAIN 197 764 PROTECTIVE ANTIGEN, PA-63. FT DOMAIN 30 287 DOMAIN 1, CALCIUM-BINDING. FT DOMAIN 288 516 DOMAIN 2, MEMBRANE INSERTION AND FT HEPTAMERIZATION. FT DOMAIN 517 624 DOMAIN 3, HEPTAMERIZATION. FT DOMAIN 625 764 DOMAIN 4, BINDING TO THE RECEPTOR; LF AND FT EF BINDING SITES. FT CA_BIND 206 206 FT CA_BIND 208 208 FT CA_BIND 210 210 FT CA_BIND 217 217 FT SITE 196 197 CLEAVAGE BY FURIN. FT SITE 343 344 CHYMOTRYPSIN CLEAVAGE; REQUIRED FOR FT TRANSLOCATION OF LF AND EF. FT VARIANT 560 560 F -> L (IN SVERDLOVSK SAMPLE). FT VARIANT 565 565 P -> S (IN STRAIN BA1024). FT VARIANT 600 600 A -> V (IN STRAINS BA1024 AND V770-NP1- FT R). FT MUTAGEN 289 289 P->A: REDUCED TOXICITY IN COMBINATION FT WITH LETHAL FACTOR; DECREASED MEMBRANE FT INSERTION AND TRANSLOCATION OF THE LETHAL FT FACTOR. FT MUTAGEN 342 342 F->C: LOSS OF TOXICITY PROBABLY DUE TO FT LOSS OF CAPABILITY TO TRANSLOCATE LF. FT MUTAGEN 342 344 FFD->AAA: DECREASE IN TOXICITY PROBABLY FT DUE TO SLOW TRANSLOCATION OF LF. FT MUTAGEN 342 343 MISSING: LOSS OF TOXICITY PROBABLY DUE TO FT LOSS OF CAPABILITY TO TRANSLOCATE LF. FT MUTAGEN 344 344 D->A: DECREASE IN TOXICITY PROBABLY DUE FT TO SLOW TRANSLOCATION OF LF. FT MUTAGEN 375 375 W->A: LOSS OF TOXICITY PROBABLY DUE TO FT FAULTY MEMBRANE INSERTION OR FT TRANSLOCATION OF LF/EF INTO THE CYTOSOL. FT MUTAGEN 379 379 M->A: NO EFFECT. FT MUTAGEN 381 381 L->A: LOSS OF TOXICITY PROBABLY DUE TO FT FAULTY MEMBRANE INSERTION OR FT TRANSLOCATION OF LF/EF INTO THE CYTOSOL. FT MUTAGEN 426 426 K->A: LOSS OF CAPABILITY TO UNDERGO FT CONFORMATIONAL CHANGES THAT LEAD TO PORE FT FORMATION AND TRANSLOCATION. FT MUTAGEN 454 454 D->A: LOSS OF CAPABILITY TO UNDERGO FT CONFORMATIONAL CHANGES THAT LEAD TO PORE FT FORMATION AND TRANSLOCATION. FT MUTAGEN 456 456 F->A: LOSS OF CAPABILITY TO UNDERGO FT CONFORMATIONAL CHANGES THAT LEAD TO PORE FT FORMATION AND TRANSLOCATION. FT MUTAGEN 512 512 Q->A: LOSS OF HEPTAMERIZATION CAPABILITY. FT MUTAGEN 541 541 D->A: LOSS OF HEPTAMERIZATION CAPABILITY. FT MUTAGEN 543 543 L->A: DECREASE IN HEPTAMERIZATION FT CAPABILITY. FT MUTAGEN 581 581 F->A: LOSS OF TOXICITY DUE TO DEFECTIVE FT OLIGOMERIZATION. FT MUTAGEN 583 583 F->A: DECREASE IN TOXICITY DUE TO FT DEFECTIVE OLIGOMERIZATION. FT MUTAGEN 591 591 I->A: LOSS OF TOXICITY DUE TO DEFECTIVE FT OLIGOMERIZATION. FT MUTAGEN 595 595 L->A: LOSS OF TOXICITY DUE TO DEFECTIVE FT OLIGOMERIZATION. FT MUTAGEN 603 603 I->A: LOSS OF TOXICITY DUE TO DEFECTIVE FT OLIGOMERIZATION. FT MUTAGEN 621 621 R->A: NO EFFECT. FT CONFLICT 314 314 Q -> E (IN REF. 1). SQ SEQUENCE 764 AA; 85810 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 //