Skip to main content
SpringerLink
Log in

Anthrax Toxin Protective Antigen Forms an Unusual Channel That Unfolds and Translocates Proteins Across Membranes

  • Chapter
Electrophysiology of Unconventional Channels and Pores

Part of the book series: Springer Series in Biophysics ((BIOPHYSICS,volume 18))

Abstract

Anthrax toxin is one of two major virulence factors secreted by pathogenic Bacillus anthracis, the etiologic agent of anthrax. Because inhalational anthrax is highly fatal, the agent has been weaponized for biowarfare and bioterrorism. Anthrax toxin is comprised of three individually nontoxic proteins, protective antigen (PA), lethal factor (LF), and edema factor (EF). But, to physiologically function, these individual subunits assemble into potent cytotoxins containing PA plus LF and/or EF. The PA component oligomerizes into a heptamer or octamer, which can insert into a host cell membrane to form a protein translocase channel. Under a transmembrane proton gradient driving force, LF and EF translocate through the narrow PA channel into the cytosol of the host cell. The narrowness of the channel necessitates that LF and EF unfold during translocation. This channel is unusual in this respect, because it contains its own unfoldase and translocase machinery. Highly nonspecific and dynamic clamp sites in the PA channel catalyze these activities. Anthrax toxin has been used extensively as a biophysical model to interrogate the molecular basis of translocation-coupled unfolding and translocation. It is being actively targeted by therapeutics to inhibit its function. New biotechnological adaptations use the toxin as a cancer therapy and generalized protein delivery vehicle.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  • Arora N, Leppla SH (1993) Residues 1-254 of anthrax toxin lethal factor are sufficient to cause cellular uptake of fused polypeptides. J Biol Chem 268:3334–3341

    CAS  PubMed  Google Scholar 

  • Ashiuchi M, Nawa C, Kamei T et al (2001) Physiological and biochemical characteristics of poly gamma-glutamate synthetase complex of Bacillus subtilis. Eur J Biochem FEBS 268:5321–5328

    CAS  Google Scholar 

  • Astumian RD (1997) Thermodynamics and kinetics of a Brownian motor. Science 276:917–922

    CAS  PubMed  Google Scholar 

  • Aubin-Tam ME, Olivares AO, Sauer RT et al (2011) Single-molecule protein unfolding and translocation by an ATP-fueled proteolytic machine. Cell 145:257–267. doi:10.1016/j.cell.2011.03.036, S0092-8674(11)00313-8 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Baldari CT, Tonello F, Paccani SR, Montecucco C (2006) Anthrax toxins: a paradigm of bacterial immune suppression. Trends Immunol 27:434–440. doi:10.1016/j.it.2006.07.002

    Article  CAS  PubMed  Google Scholar 

  • Bann JG (2012) Anthrax toxin protective antigen-Insights into molecular switching from prepore to pore. Protein Sci 21:1–12. doi:10.1002/pro.752

    Article  CAS  PubMed  Google Scholar 

  • Barth H, Aktories K, Popoff MR, Stiles BG (2004) Binary bacterial toxins: biochemistry, biology, and applications of common Clostridium and Bacillus proteins. Microbiol Mol Biol Rev 68:373–402

    CAS  PubMed  PubMed Central  Google Scholar 

  • Basilio D, Jennings-Antipov LD, Jakes KS, Finkelstein A (2011a) Trapping a translocating protein within the anthrax toxin channel: implications for the secondary structure of permeating proteins. J Gen Physiol 137:343–356. doi:10.1085/jgp.201010578, jgp.201010578 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Basilio D, Juris SJ, Collier RJ, Finkelstein A (2009) Evidence for a proton-protein symport mechanism in the anthrax toxin channel. J Gen Physiol 133:307–314

    CAS  PubMed  PubMed Central  Google Scholar 

  • Basilio D, Kienker PK, Briggs SW, Finkelstein A (2011b) A kinetic analysis of protein transport through the anthrax toxin channel. J Gen Physiol 137:521–531. doi:10.1085/jgp.201110627, jgp.201110627 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bell SE, Mavila A, Salazar R et al (2001) Differential gene expression during capillary morphogenesis in 3D collagen matrices: regulated expression of genes involved in basement membrane matrix assembly, cell cycle progression, cellular differentiation and G-protein signaling. J Cell Sci 114:2755–2773

    CAS  PubMed  Google Scholar 

  • Belton FC, Strange RE (1954) Studies on a protective antigen produced in vitro from Bacillus anthracis: medium and methods of production. Br J Exp Pathol 35:144–152

    CAS  PubMed  PubMed Central  Google Scholar 

  • Benson EL, Huynh PD, Finkelstein A, Collier RJ (1998) Identification of residues lining the anthrax protective antigen channel. Biochemistry 37:3941–3948

    CAS  PubMed  Google Scholar 

  • Beyer W, Turnbull PCB (2009) Anthrax in animals. Mol Aspects Med 30:481–489. doi:10.1016/j.mam.2009.08.004

    Article  CAS  PubMed  Google Scholar 

  • Bezrukov SM, Liu X, Karginov VA et al (2012) Interactions of high-affinity cationic blockers with the translocation pores of B. anthracis, C. botulinum, and C. perfringens binary toxins. Biophys J 103:1208–1217. doi:10.1016/j.bpj.2012.07.050

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Blanke SR, Milne JC, Benson EL, Collier RJ (1996) Fused polycationic peptide mediates delivery of diphtheria toxin A chain to the cytosol in the presence of anthrax protective antigen. Proc Natl Acad Sci U S A 93:8437–8442

    CAS  PubMed  PubMed Central  Google Scholar 

  • Blaustein RO, Finkelstein A (1990) Voltage-dependent block of anthrax toxin channels in planar phospholipid bilayer membranes by symmetric tetraalkylammonium ions. Effects on macroscopic conductance. J Gen Physiol 96:905–919

    CAS  PubMed  Google Scholar 

  • Blaustein RO, Koehler TM, Collier RJ, Finkelstein A (1989) Anthrax toxin: channel-forming activity of protective antigen in planar phospholipid bilayers. Proc Natl Acad Sci U S A 86:2209–2213

    CAS  PubMed  PubMed Central  Google Scholar 

  • Blaustein RO, Lea EJ, Finkelstein A (1990) Voltage-dependent block of anthrax toxin channels in planar phospholipid bilayer membranes by symmetric tetraalkylammonium ions. Single-channel analysis. J Gen Physiol 96:921–942

    CAS  PubMed  Google Scholar 

  • Boyer AE, Quinn CP, Hoffmaster AR et al (2009) Kinetics of lethal factor and poly-d-glutamic acid antigenemia during inhalation anthrax in rhesus macaques. Infect Immun 77:3432–3441. doi:10.1128/IAI.00346-09, IAI.00346-09 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bradley KA, Mogridge J, Mourez M et al (2001) Identification of the cellular receptor for anthrax toxin. Nature 414:225–229

    CAS  PubMed  Google Scholar 

  • Brown MJ, Thoren KL, Krantz BA (2011) Charge requirements for proton gradient-driven translocation of anthrax toxin. J Biol Chem 286:23189–23199. doi:10.1074/jbc.M111.231167, M111.231167 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bruckner V, Kovacs J, Denes G (1953) Structure of poly-d-glutamic acid isolated from capsulated strains of B. anthracis. Nature 172:508

    CAS  PubMed  Google Scholar 

  • Burton RE, Siddiqui SM, Kim YI et al (2001) Effects of protein stability and structure on substrate processing by the ClpXP unfolding and degradation machine. EMBO J 20:3092–3100

    CAS  PubMed  PubMed Central  Google Scholar 

  • Candela T, Fouet A (2005) Bacillus anthracis CapD, belonging to the γ-glutamyltranspeptidase family, is required for the covalent anchoring of capsule to peptidoglycan. Mol Microbiol 57:717–726

    CAS  PubMed  Google Scholar 

  • Candela T, Fouet A (2006) Poly-gamma-glutamate in bacteria. Mol Microbiol 60:1091–1098. doi:10.1111/j.1365-2958.2006.05179.x

    Article  CAS  PubMed  Google Scholar 

  • Candela T, Mock M, Fouet A (2005) CapE, a 47-amino-acid peptide, is necessary for Bacillus anthracis polyglutamate capsule synthesis. J Bacteriol 187:7765–7772. doi:10.1128/JB.187.22.7765-7772.2005

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chauwin JF, Oster G, Glick BS (1998) Strong precursor-pore interactions constrain models for mitochondrial protein import. Biophys J 74:1732–1743

    CAS  PubMed  PubMed Central  Google Scholar 

  • Chavarría-Smith J, Vance RE (2013) Direct proteolytic cleavage of NLRP1B is necessary and sufficient for inflammasome activation by anthrax lethal factor. PLoS Pathog 9, e1003452. doi:10.1371/journal.ppat.1003452

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Christensen KA, Krantz BA, Melnyk RA, Collier RJ (2005) Interaction of the 20 kDa and 63 kDa fragments of anthrax protective antigen: kinetics and thermodynamics. Biochemistry 44:1047–1053. doi:10.1021/bi047791s

    Article  CAS  PubMed  Google Scholar 

  • Collier RJ (2009) Membrane translocation by anthrax toxin. Mol Aspects Med 30:413–422. doi:10.1016/j.mam.2009.06.003, S0098-2997(09)00035-1 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Collier RJ, Young JA (2003) Anthrax toxin. Annu Rev Cell Dev Biol 19:45–70

    CAS  PubMed  Google Scholar 

  • Crampton N, Brockwell DJ (2010) Unravelling the design principles for single protein mechanical strength. Curr Opin Struct Biol 20:508–517. doi:10.1016/j.sbi.2010.05.005, S0959-440X(10)00075-8 [pii]

    Article  CAS  PubMed  Google Scholar 

  • Cunningham K, Lacy DB, Mogridge J, Collier RJ (2002) Mapping the lethal factor and edema factor binding sites on oligomeric anthrax protective antigen. Proc Natl Acad Sci U S A 99:7049–7053

    CAS  PubMed  PubMed Central  Google Scholar 

  • Gill DM (1978) Seven toxic peptides that cross cell membranes. In: Jeljaszewicz J, Wadstrom T (eds) Bacterial Toxins and Cell Membranes. Academic, New York, pp 291–332

    Google Scholar 

  • Dmochewitz L, Lillich M, Kaiser E et al (2011) Role of CypA and Hsp90 in membrane translocation mediated by anthrax protective antigen. Cell Microbiol 13:359–373. doi:10.1111/j.1462-5822.2010.01539.x

    Article  CAS  PubMed  Google Scholar 

  • Drum CL, Yan SZ, Bard J et al (2002) Structural basis for the activation of anthrax adenylyl cyclase exotoxin by calmodulin. Nature 415:396–402

    CAS  PubMed  Google Scholar 

  • Duesbery NS, Webb CP, Leppla SH et al (1998) Proteolytic inactivation of MAP-kinase-kinase by anthrax lethal factor. Science 280:734–737

    CAS  PubMed  Google Scholar 

  • Dumetz F, Jouvion G, Khun H et al (2011) Noninvasive imaging technologies reveal edema toxin as a key virulence factor in anthrax. Am J Pathol 178:2523–2535. doi:10.1016/j.ajpath.2011.02.027

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Evans AS (1976) Causation and disease: the Henle-Koch postulates revisited. Yale J Biol Med 49:175–195

    CAS  PubMed  PubMed Central  Google Scholar 

  • Ezzell JW, Abshire TG (1992) Serum protease cleavage of Bacillus anthracis protective antigen. J Gen Microbiol 138:543–549

    CAS  PubMed  Google Scholar 

  • Ezzell JW, Abshire TG, Panchal R et al (2009) Association of Bacillus anthracis capsule with lethal toxin during experimental infection. Infect Immun 77:749–755. doi:10.1128/IAI.00764-08, IAI.00764-08 [pii]

    Article  CAS  PubMed  Google Scholar 

  • Feld GK, Brown MJ, Krantz BA (2012a) Ratcheting up protein translocation with anthrax toxin. Protein Sci 21:606–624. doi:10.1002/pro.2052

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Feld GK, Kintzer AF, Tang II et al (2012b) Domain flexibility modulates the heterogeneous assembly mechanism of anthrax toxin protective antigen. J Mol Biol 415:159–174. doi:10.1016/j.jmb.2011.10.035, S0022-2836(11)01172-7 [pii]

    Article  CAS  PubMed  Google Scholar 

  • Feld GK, Thoren KL, Kintzer AF et al (2010) Structural basis for the unfolding of anthrax lethal factor by protective antigen oligomers. Nat Struct Mol Biol 17:1383–1390. doi:10.1038/nsmb.1923, nsmb.1923 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Feller W (2008) An introduction to probability theory and its applications. Wiley, Hoboken

    Google Scholar 

  • Fersht AR (1998) Structure and mechanism in protein science: a guide to enzyme catalysis and protein folding. Freeman, New York

    Google Scholar 

  • Finkelstein A (2009) Proton-coupled protein transport through the anthrax toxin channel. Philos Trans R Soc Lond B Biol Sci 364:209–215. doi:10.1098/rstb.2008.0126, 74702356P024100L [pii]

    Article  CAS  PubMed  Google Scholar 

  • Floyd DL, Ragains JR, Skehel JJ et al (2008) Single-particle kinetics of influenza virus membrane fusion. Proc Natl Acad Sci U S A 105:15382–15387. doi:10.1073/pnas.0807771105

    Article  PubMed  PubMed Central  Google Scholar 

  • Frankel AE, Kuo S-R, Dostal D et al (2009) Pathophysiology of anthrax. Front Biosci Landmark Ed 14:4516–4524

    CAS  PubMed  PubMed Central  Google Scholar 

  • Friedlander AM (1986) Macrophages are sensitive to anthrax lethal toxin through an acid-dependent process. J Biol Chem 261:7123–7126

    CAS  PubMed  Google Scholar 

  • Friedlander AM, Pittman PR, Parker GW (1999) Anthrax vaccine: evidence for safety and efficacy against inhalational anthrax. JAMA 282:2104–2106

    CAS  PubMed  Google Scholar 

  • Fu S, Tong X, Cai C et al (2010) The structure of tumor endothelial marker 8 (TEM8) extracellular domain and implications for its receptor function for recognizing anthrax toxin. PLoS One 5, e11203. doi:10.1371/journal.pone.0011203

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Glick BS (1995) Can Hsp70 proteins act as force-generating motors? Cell 80:11–14

    CAS  PubMed  Google Scholar 

  • Green BD, Battisti L, Koehler TM et al (1985) Demonstration of a capsule plasmid in Bacillus anthracis. Infect Immun 49:291–297

    CAS  PubMed  PubMed Central  Google Scholar 

  • Gu LQ, Braha O, Conlan S et al (1999) Stochastic sensing of organic analytes by a pore-forming protein containing a molecular adapter. Nature 398:686–690

    CAS  Google Scholar 

  • Halverson KM, Panchal RG, Nguyen TL et al (2005) Anthrax biosensor: protective antigen ion channel asymmetric blockade. J Biol Chem 280:34056–34062

    CAS  PubMed  Google Scholar 

  • Hanks S, Adams S, Douglas J et al (2003) Mutations in the gene encoding capillary morphogenesis protein 2 cause juvenile hyaline fibromatosis and infantile systemic hyalinosis. Am J Hum Genet 73:791–800

    CAS  PubMed  PubMed Central  Google Scholar 

  • Huang S, Ratliff KS, Schwartz MP et al (1999) Mitochondria unfold precursor proteins by unraveling them from their N-termini. Nat Struct Biol 6:1132–1138

    CAS  PubMed  Google Scholar 

  • Jang J, Cho M, Chun JH et al (2011) The poly-γ-d-glutamic acid capsule of Bacillus anthracis enhances lethal toxin activity. Infect Immun 79:3846–3854. doi:10.1128/IAI.01145-10, IAI.01145-10 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Janowiak BE, Finkelstein A, Collier RJ (2009) An approach to characterizing single-subunit mutations in multimeric prepores and pores of anthrax protective antigen. Protein Sci 18:348–358. doi:10.1002/pro.35

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jernigan DB, Raghunathan PL, Bell BP et al (2002) Investigation of bioterrorism-related anthrax, United States, 2001: epidemiologic findings. Emerg Infect Dis 8:1019–1028. doi:10.3201/eid0810.020353

    Article  PubMed  PubMed Central  Google Scholar 

  • Juris SJ, Melnyk RA, Bolcome RE et al (2007) Cross-linked forms of the isolated N-terminal domain of the lethal factor are potent inhibitors of anthrax toxin. Infect Immun 75:5052–5058. doi:10.1128/IAI.00490-07, IAI.00490-07 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Karginov VA, Nestorovich EM, Moayeri M et al (2005) Blocking anthrax lethal toxin at the protective antigen channel by using structure-inspired drug design. Proc Natl Acad Sci U S A 102:15075–15080. doi:10.1073/pnas.0507488102

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Katayama H, Janowiak BE, Brzozowski M et al (2008) GroEL as a molecular scaffold for structural analysis of the anthrax toxin pore. Nat Struct Mol Biol 15:754–760. doi:10.1038/nsmb.1442, nsmb.1442 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Katayama H, Wang J, Tama F et al (2010) Three-dimensional structure of the anthrax toxin pore inserted into lipid nanodiscs and lipid vesicles. Proc Natl Acad Sci U S A 107:3453–3457

    CAS  PubMed  PubMed Central  Google Scholar 

  • Kenniston JA, Baker TA, Fernandez JM, Sauer RT (2003) Linkage between ATP consumption and mechanical unfolding during the protein processing reactions of an AAA+ degradation machine. Cell 114:511–520

    CAS  PubMed  Google Scholar 

  • Kintzer AF, Sterling HJ, Tang II et al (2010a) Anthrax toxin receptor drives protective antigen oligomerization and stabilizes the heptameric and octameric oligomer by a similar mechanism. PLoS One 5, e13888

    PubMed  PubMed Central  Google Scholar 

  • Kintzer AF, Sterling HJ, Tang II et al (2010b) Role of the protective antigen octamer in the molecular mechanism of anthrax lethal toxin stabilization in plasma. J Mol Biol 399:741–758. doi:10.1016/j.jmb.2010.04.041, S0022-2836(10)00429-8 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kintzer AF, Tang II, Schawel AK et al (2012) Anthrax toxin protective antigen integrates poly-γ-d-glutamate and pH signals to sense the optimal environment for channel formation. Proc Natl Acad Sci U S A 109:18378–18383. doi:10.1073/pnas.1208280109, 1208280109 [pii]

    Article  PubMed  PubMed Central  Google Scholar 

  • Kintzer AF, Thoren KL, Sterling HJ et al (2009) The protective antigen component of anthrax toxin forms functional octameric complexes. J Mol Biol 392:614–629. doi:10.1016/j.jmb.2009.07.037, S0022-2836(09)00876-6 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Klimpel KR, Molloy SS, Thomas G, Leppla SH (1992) Anthrax toxin protective antigen is activated by a cell surface protease with the sequence specificity and catalytic properties of furin. Proc Natl Acad Sci U S A 89:10277–10281

    CAS  PubMed  PubMed Central  Google Scholar 

  • Koehler TM (2009) Bacillus anthracis physiology and genetics. Mol Aspects Med 30:386–396. doi:10.1016/j.mam.2009.07.004, S0098-2997(09)00052-1 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kofoed EM, Vance RE (2011) Innate immune recognition of bacterial ligands by NAIPs determines inflammasome specificity. Nature 477:592–595. doi:10.1038/nature10394, nature10394 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Krantz BA, Finkelstein A, Collier RJ (2006) Protein translocation through the anthrax toxin transmembrane pore is driven by a proton gradient. J Mol Biol 355:968–979

    CAS  PubMed  Google Scholar 

  • Krantz BA, Melnyk RA, Zhang S et al (2005) A phenylalanine clamp catalyzes protein translocation through the anthrax toxin pore. Science 309:777–781

    CAS  PubMed  PubMed Central  Google Scholar 

  • Krantz BA, Trivedi AD, Cunningham K et al (2004) Acid-induced unfolding of the amino-terminal domains of the lethal and edema factors of anthrax toxin. J Mol Biol 344:739–756

    CAS  PubMed  Google Scholar 

  • Lacy DB, Lin HC, Melnyk RA et al (2005) A model of anthrax toxin lethal factor bound to protective antigen. Proc Natl Acad Sci U S A 102:16409–16414

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lacy DB, Mourez M, Fouassier A, Collier RJ (2002) Mapping the anthrax protective antigen binding site on the lethal and edema factors. J Biol Chem 277:3006–3010

    CAS  PubMed  Google Scholar 

  • Lacy DB, Wigelsworth DJ, Melnyk RA et al (2004a) Structure of heptameric protective antigen bound to an anthrax toxin receptor: a role for receptor in pH-dependent pore formation. Proc Natl Acad Sci U S A 101:13147–13151

    CAS  PubMed  PubMed Central  Google Scholar 

  • Lacy DB, Wigelsworth DJ, Scobie HM et al (2004b) Crystal structure of the von Willebrand factor A domain of human capillary morphogenesis protein 2: an anthrax toxin receptor. Proc Natl Acad Sci U S A 101:6367–6372. doi:10.1073/pnas.0401506101 [doi] 0401506101 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Leppla SH (1982) Anthrax toxin edema factor: a bacterial adenylate cyclase that increases cyclic AMP concentrations of eukaryotic cells. Proc Natl Acad Sci U S A 79:3162–3166

    CAS  PubMed  PubMed Central  Google Scholar 

  • Levinsohn JL, Newman ZL, Hellmich KA et al (2012) Anthrax lethal factor cleavage of Nlrp1 is required for activation of the inflammasome. PLoS Pathog 8:e1002638. doi:10.1371/journal.ppat.1002638, PPATHOGENS-D-12-00046 [pii]

    Article  PubMed  PubMed Central  Google Scholar 

  • Liao X, Rabideau AE, Pentelute BL (2014) Delivery of antibody mimics into mammalian cells via anthrax toxin protective antigen. Chembiochem 15:2458–2466. doi:10.1002/cbic.201402290

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Liu S, Crown D, Miller-Randolph S et al (2009) Capillary morphogenesis protein-2 is the major receptor mediating lethality of anthrax toxin in vivo. Proc Natl Acad Sci U S A 106:12424–12429. doi:10.1073/pnas.0905409106

    Article  PubMed  PubMed Central  Google Scholar 

  • Maillard RA, Chistol G, Sen M et al (2011) ClpX(P) generates mechanical force to unfold and translocate its protein substrates. Cell 145:459–469. doi:10.1016/j.cell.2011.04.010, S0092-8674(11)00429-6 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Martchenko M, Jeong S-Y, Cohen SN (2010) Heterodimeric integrin complexes containing beta1-integrin promote internalization and lethality of anthrax toxin. Proc Natl Acad Sci U S A 107:15583–15588. doi:10.1073/pnas.1010145107

    Article  PubMed  PubMed Central  Google Scholar 

  • Mayor A (2008) Greek fire, poison arrows, and scorpion bombs: biological & chemical warfare in the ancient world. Overlook Press, New York

    Google Scholar 

  • McCluskey AJ, Collier RJ (2013) Receptor-directed chimeric toxins created by sortase-mediated protein fusion. Mol Cancer Ther 12:2273–2281. doi:10.1158/1535-7163.MCT-13-0358

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Meador WE, Means AR, Quiocho FA (1992) Target enzyme recognition by calmodulin: 2.4 a structure of a calmodulin-peptide complex. Science 257:1251–1255

    CAS  PubMed  Google Scholar 

  • Meador WE, Means AR, Quiocho FA (1993) Modulation of calmodulin plasticity in molecular recognition on the basis of x-ray structures. Science 262:1718–1721

    CAS  PubMed  Google Scholar 

  • Melnyk RA, Collier RJ (2006) A loop network within the anthrax toxin pore positions the phenylalanine clamp in an active conformation. Proc Natl Acad Sci U S A 103:9802–9807. doi:10.1073/pnas.0604000103, 0604000103 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Melnyk RA, Hewitt KM, Lacy DB et al (2006) Structural determinants for the binding of anthrax lethal factor to oligomeric protective antigen. J Biol Chem 281:1630–1635

    CAS  PubMed  Google Scholar 

  • Mikesell P, Ivins BE, Ristroph JD, Dreier TM (1983) Evidence for plasmid-mediated toxin production in Bacillus anthracis. Infect Immun 39:371–376

    CAS  PubMed  PubMed Central  Google Scholar 

  • Miller CJ, Elliott JL, Collier RJ (1999) Anthrax protective antigen: prepore-to-pore conversion. Biochemistry 38:10432–10441

    CAS  PubMed  Google Scholar 

  • Milne JC, Blanke SR, Hanna PC, Collier RJ (1995) Protective antigen-binding domain of anthrax lethal factor mediates translocation of a heterologous protein fused to its amino- or carboxy-terminus. Mol Microbiol 15:661–666

    CAS  PubMed  Google Scholar 

  • Milne JC, Furlong D, Hanna PC et al (1994) Anthrax protective antigen forms oligomers during intoxication of mammalian cells. J Biol Chem 269:20607–20612

    CAS  PubMed  Google Scholar 

  • Moayeri M, Leppla SH (2009) Cellular and systemic effects of anthrax lethal toxin and edema toxin. Mol Aspects Med 30:439–455. doi:10.1016/j.mam.2009.07.003, S0098-2997(09)00051-X [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Moayeri M, Wiggins JF, Leppla SH (2007) Anthrax protective antigen cleavage and clearance from the blood of mice and rats. Infect Immun 75:5175–5184. doi:10.1128/IAI.00719-07, IAI.00719-07 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mock M, Fouet A (2001) Anthrax. Annu Rev Microbiol 55:647–671

    CAS  PubMed  Google Scholar 

  • Mogridge J, Cunningham K, Collier RJ (2002a) Stoichiometry of anthrax toxin complexes. Biochemistry 41:1079–1082

    CAS  PubMed  Google Scholar 

  • Mogridge J, Cunningham K, Lacy DB et al (2002b) The lethal and edema factors of anthrax toxin bind only to oligomeric forms of the protective antigen. Proc Natl Acad Sci U S A 99:7045–7048

    CAS  PubMed  PubMed Central  Google Scholar 

  • Mogridge J, Mourez M, Collier RJ (2001) Involvement of domain 3 in oligomerization by the protective antigen moiety of anthrax toxin. J Bacteriol 183:2111–2116

    CAS  PubMed  PubMed Central  Google Scholar 

  • Molloy SS, Bresnahan PA, Leppla SH et al (1992) Human furin is a calcium-dependent serine endoprotease that recognizes the sequence Arg-X-X-Arg and efficiently cleaves anthrax toxin protective antigen. J Biol Chem 267:16396–16402

    CAS  PubMed  Google Scholar 

  • Mourez M, Yan M, Lacy DB et al (2003) Mapping dominant-negative mutations of anthrax protective antigen by scanning mutagenesis. Proc Natl Acad Sci U S A 100:13803–13808

    CAS  PubMed  PubMed Central  Google Scholar 

  • Mueller P, Rudin DO, Tien HT, Westcott WC (1963) Methods for the formation of single bimolecular lipid membranes in aqueous solution. J Phys Chem 67:534–535

    CAS  Google Scholar 

  • Nanda A, Carson-Walter EB, Seaman S et al (2004) TEM8 interacts with the cleaved C5 domain of collagen alpha 3(VI). Cancer Res 64:817–820

    CAS  PubMed  Google Scholar 

  • Nassi S, Collier RJ, Finkelstein A (2002) PA63 channel of anthrax toxin: an extended β-barrel. Biochemistry 41:1445–1450

    CAS  PubMed  Google Scholar 

  • Nestorovich EM, Bezrukov SM (2014) Designing inhibitors of anthrax toxin. Expert Opin Drug Discov 9:299–318. doi:10.1517/17460441.2014.877884

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Nguyen TL (2004) Three-dimensional model of the pore form of anthrax protective antigen. Structure and biological implications. J Biomol Struct Dyn 22:253–265

    CAS  PubMed  Google Scholar 

  • Oomen CJ, Van Ulsen P, Van Gelder P et al (2004) Structure of the translocator domain of a bacterial autotransporter. EMBO J 23:1257–1266

    CAS  PubMed  PubMed Central  Google Scholar 

  • Orlik F, Schiffler B, Benz R (2005) Anthrax toxin protective antigen: inhibition of channel function by chloroquine and related compounds and study of binding kinetics using the current noise analysis. Biophys J 88:1715–1724

    CAS  PubMed  Google Scholar 

  • Panchal RG, Halverson KM, Ribot W et al (2005) Purified Bacillus anthracis lethal toxin complex formed in vitro and during infection exhibits functional and biological activity. J Biol Chem 280:10834–10839. doi:10.1074/jbc.M412210200

    Article  CAS  PubMed  Google Scholar 

  • Pannifer AD, Wong TY, Schwarzenbacher R et al (2001) Crystal structure of the anthrax lethal factor. Nature 414:229–233

    CAS  PubMed  Google Scholar 

  • Pentelute BL, Barker AP, Janowiak BE et al (2010) A semisynthesis platform for investigating structure-function relationships in the N-terminal domain of the anthrax Lethal Factor. ACS Chem Biol 5:359–364. doi:10.1021/cb100003r

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pentelute BL, Sharma O, Collier RJ (2011) Chemical dissection of protein translocation through the anthrax toxin pore. Angew Chem Int Ed Engl 50:2294–2296. doi:10.1002/anie.201006460

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Petosa C, Collier RJ, Klimpel KR et al (1997) Crystal structure of the anthrax toxin protective antigen. Nature 385:833–838

    CAS  PubMed  Google Scholar 

  • Pflughoeft KJ, Swick MC, Engler DA et al (2014) Modulation of the Bacillus anthracis secretome by the immune inhibitor A1 protease. J Bacteriol 196:424–435. doi:10.1128/JB.00690-13

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Phillips DD, Fattah RJ, Crown D et al (2013) Engineering anthrax toxin variants that exclusively form octamers and their application to targeting tumors. J Biol Chem 288:9058–9065. doi:10.1074/jbc.M113.452110

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pittman PR, Kim-Ahn G, Pifat DY et al (2002) Anthrax vaccine: immunogenicity and safety of a dose-reduction, route-change comparison study in humans. Vaccine 20:1412–1420

    CAS  PubMed  Google Scholar 

  • Quinn CP, Singh Y, Klimpel KR, Leppla SH (1991) Functional mapping of anthrax toxin lethal factor by in-frame insertion mutagenesis. J Biol Chem 266:20124–20130

    CAS  PubMed  Google Scholar 

  • Rasko DA, Worsham PL, Abshire TG et al (2011) Bacillus anthracis comparative genome analysis in support of the Amerithrax investigation. Proc Natl Acad Sci U S A 108:5027–5032. doi:10.1073/pnas.1016657108, 1016657108 [pii]

    Article  PubMed  PubMed Central  Google Scholar 

  • Ratts R, Trujillo C, Bharti A et al (2005) A conserved motif in transmembrane helix 1 of diphtheria toxin mediates catalytic domain delivery to the cytosol. Proc Natl Acad Sci U S A 102:15635–15640. doi:10.1073/pnas.0504937102, 0504937102 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ratts R, Zeng H, Berg EA et al (2003) The cytosolic entry of diphtheria toxin catalytic domain requires a host cell cytosolic translocation factor complex. J Cell Biol 160:1139–1150. doi:10.1083/jcb.200210028, jcb.200210028 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ryan PL, Young JA (2008) Evidence against a human cell-specific role for LRP6 in anthrax toxin entry. PLoS One 3:e1817. doi:10.1371/journal.pone.0001817

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Santelli E, Bankston LA, Leppla SH, Liddington RC (2004) Crystal structure of a complex between anthrax toxin and its host cell receptor. Nature 430:905–908

    CAS  PubMed  Google Scholar 

  • Schueler-Furman O, Wang C, Baker D (2005) Progress in protein-protein docking: atomic resolution predictions in the CAPRI experiment using RosettaDock with an improved treatment of side-chain flexibility. Proteins 60:187–194. doi:10.1002/prot.20556

    Article  CAS  PubMed  Google Scholar 

  • Scobie HM, Rainey GJ, Bradley KA, Young JA (2003) Human capillary morphogenesis protein 2 functions as an anthrax toxin receptor. Proc Natl Acad Sci U S A 100:5170–5174. doi:10.1073/pnas.0431098100, 0431098100 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sellman BR, Mourez M, Collier RJ (2001a) Dominant-negative mutants of a toxin subunit: an approach to therapy of anthrax. Science 292:695–697

    CAS  PubMed  Google Scholar 

  • Sellman BR, Nassi S, Collier RJ (2001b) Point mutations in anthrax protective antigen that block translocation. J Biol Chem 276:8371–8376

    CAS  PubMed  Google Scholar 

  • Sharma O, Collier RJ (2014) Polylysine-mediated translocation of the diphtheria toxin catalytic domain through the anthrax protective antigen pore. Biochemistry 53:6934–6940. doi:10.1021/bi500985v

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Shaw CA, Starnbach MN (2008) Antigen delivered by anthrax lethal toxin induces the development of memory CD8+ T cells that can be rapidly boosted and display effector functions. Infect Immun 76:1214–1222. doi:10.1128/IAI.01208-07

    Article  CAS  PubMed  Google Scholar 

  • Shen Y, Zhukovskaya NL, Guo Q et al (2005) Calcium-independent calmodulin binding and two-metal-ion catalytic mechanism of anthrax edema factor. EMBO J 24:929–941

    CAS  PubMed  PubMed Central  Google Scholar 

  • Simon SM, Peskin CS, Oster GF (1992) What drives the translocation of proteins? Proc Natl Acad Sci U S A 89:3770–3774

    CAS  PubMed  PubMed Central  Google Scholar 

  • Singh Y, Klimpel KR, Goel S et al (1999) Oligomerization of anthrax toxin protective antigen and binding of lethal factor during endocytic uptake into mammalian cells. Infect Immun 67:1853–1859

    CAS  PubMed  PubMed Central  Google Scholar 

  • Smith H, Keppie J (1954) Observations on experimental anthrax: demonstration of a specific lethal factor produced in vivo by Bacillus anthracis. Nature 173:689

    Google Scholar 

  • Song L, Hobaugh MR, Shustak C et al (1996) Structure of staphylococcal α-hemolysin, a heptameric transmembrane pore. Science 274:1859–1866

    CAS  PubMed  Google Scholar 

  • Sterne M (1939) The use of anthrax vaccines prepared from avirulent (uncapsulated) variants of Bacillus anthracis. Ondersteport J Vet Sci Anim Indust 13:307–312

    Google Scholar 

  • Strange RE, Belton FC (1954) Studies on a protective antigen produced in vitro from Bacillus anthracis: purification and chemistry of the antigen. Br J Exp Pathol 35:153–165

    CAS  PubMed  PubMed Central  Google Scholar 

  • Sun J, Lang AE, Aktories K, Collier RJ (2008) Phenylalanine-427 of anthrax protective antigen functions in both pore formation and protein translocation. Proc Natl Acad Sci U S A 105:4346–4351. doi:10.1073/pnas.0800701105, 0800701105 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sun J, Vernier G, Wigelsworth DJ, Collier RJ (2007) Insertion of anthrax protective antigen into liposomal membranes: effects of a receptor. J Biol Chem 282:1059–1065

    CAS  PubMed  Google Scholar 

  • Tamayo AG, Bharti A, Trujillo C et al (2008) COPI coatomer complex proteins facilitate the translocation of anthrax lethal factor across vesicular membranes in vitro. Proc Natl Acad Sci U S A 105:5254–5259. doi:10.1073/pnas.0710100105, 0710100105 [pii]

    Article  PubMed  PubMed Central  Google Scholar 

  • Thoren KL, Krantz BA (2011) The unfolding story of anthrax toxin translocation. Mol Microbiol 80:588–595. doi:10.1111/j.1365-2958.2011.07614.x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Thoren KL, Worden EJ, Yassif JM, Krantz BA (2009) Lethal factor unfolding is the most force-dependent step of anthrax toxin translocation. Proc Natl Acad Sci U S A 106:21555–21560. doi:10.1073/pnas.0905880106, 0905880106 [pii]

    Article  PubMed  PubMed Central  Google Scholar 

  • Turnbull PC (1991) Anthrax vaccines: past, present and future. Vaccine 9:533–539

    CAS  PubMed  Google Scholar 

  • Turnbull PCB (1996) Bacillus Medical Microbiology. University of Texas Medical Branch at Galveston, Galveston

    Google Scholar 

  • Uchida I, Makino S, Sasakawa C et al (1993) Identification of a novel gene, dep, associated with depolymerization of the capsular polymer in Bacillus anthracis. Mol Microbiol 9:487–496

    CAS  PubMed  Google Scholar 

  • Uchida I, Sekizaki T, Hashimoto K, Terakado N (1985) Association of the encapsulation of Bacillus anthracis with a 60 megadalton plasmid. J Gen Microbiol 131:363–367

    CAS  PubMed  Google Scholar 

  • Van der Goot G, Young JA (2009) Receptors of anthrax toxin and cell entry. Mol Aspects Med 30:406–412. doi:10.1016/j.mam.2009.08.007, S0098-2997(09)00059-4 [pii]

    Article  CAS  PubMed  Google Scholar 

  • Vitale G, Bernardi L, Napolitani G et al (2000) Susceptibility of mitogen-activated protein kinase kinase family members to proteolysis by anthrax lethal factor. Biochem J 352:739–745

    CAS  PubMed  PubMed Central  Google Scholar 

  • Von Moltke J, Trinidad NJ, Moayeri M et al (2012) Rapid induction of inflammatory lipid mediators by the inflammasome in vivo. Nature 490:107–111. doi:10.1038/nature11351

    Article  CAS  Google Scholar 

  • Wasserman GM, Grabenstein JD, Pittman PR et al (2003) Analysis of adverse events after anthrax immunization in US Army medical personnel. J Occup Environ Med Am Coll Occup Environ Med 45:222–233

    Google Scholar 

  • Wein AN, Liu S, Zhang Y et al (2013) Tumor therapy with a urokinase plasminogen activator-activated anthrax lethal toxin alone and in combination with paclitaxel. Invest New Drugs 31:206–212. doi:10.1007/s10637-012-9847-1

    Article  PubMed  Google Scholar 

  • Wei W, Lu Q, Chaudry GJ et al (2006) The LDL receptor-related protein LRP6 mediates internalization and lethality of anthrax toxin. Cell 124:1141–1154. doi:10.1016/j.cell.2005.12.045, S0092-8674(06)00199-1 [pii]

    Article  CAS  PubMed  Google Scholar 

  • Wesche J, Elliott JL, Falnes PO et al (1998) Characterization of membrane translocation by anthrax protective antigen. Biochemistry 37:15737–15746

    CAS  PubMed  Google Scholar 

  • Wigelsworth DJ, Krantz BA, Christensen KA et al (2004) Binding stoichiometry and kinetics of the interaction of a human anthrax toxin receptor, CMG2, with protective antigen. J Biol Chem 279:23349–23356

    CAS  PubMed  Google Scholar 

  • Williams P, Wallace D (1989) Unit 731: Japan’s secret biological warfare in World War II. Free Press, New York

    Google Scholar 

  • Wimalasena DS, Cramer JC, Janowiak BE et al (2007) Effect of 2-fluorohistidine labeling of the anthrax protective antigen on stability, pore formation, and translocation. Biochemistry 46:14928–14936. doi:10.1021/bi701763z

    Article  CAS  PubMed  Google Scholar 

  • Wolfe JT, Krantz BA, Rainey GJ et al (2005) Whole-cell voltage clamp measurements of anthrax toxin pore current. J Biol Chem 280:39417–39422

    CAS  PubMed  Google Scholar 

  • Wright GG, Hedberg MA, Slein JB (1954) Studies on immunity in anthrax. III. Elaboration of protective antigen in a chemically defined, non-protein medium. J Immunol 72:263–269

    CAS  PubMed  Google Scholar 

  • Wynia-Smith SL, Brown MJ, Chirichella G et al (2012) Electrostatic ratchet in the protective antigen channel promotes anthrax toxin translocation. J Biol Chem 287:43753–43764. doi:10.1074/jbc.M112.419598, M112.419598 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Young JA, Collier RJ (2007) Anthrax toxin: receptor binding, internalization, pore formation, and translocation. Annu Rev Biochem 76:243–265. doi:10.1146/annurev.biochem.75.103004.142728

    Article  CAS  PubMed  Google Scholar 

  • Young JJ, Bromberg-White JL, Zylstra C et al (2007) LRP5 and LRP6 are not required for protective antigen-mediated internalization or lethality of anthrax lethal toxin. PLoS Pathog 3, e27. doi:10.1371/journal.ppat.0030027, 07-PLPA-RA-0017 [pii]

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang S, Finkelstein A, Collier RJ (2004a) Evidence that translocation of anthrax toxin’s lethal factor is initiated by entry of its N terminus into the protective antigen channel. Proc Natl Acad Sci U S A 101:16756–16761

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang S, Udho E, Wu Z et al (2004b) Protein translocation through anthrax toxin channels formed in planar lipid bilayers. Biophys J 87:3842–3849

    CAS  PubMed  PubMed Central  Google Scholar 

  • Zhao Y, Yang J, Shi J et al (2011) The NLRC4 inflammasome receptors for bacterial flagellin and type III secretion apparatus. Nature 477:596–600. doi:10.1038/nature10510

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Microbial Pathogenesis, School of Dentistry, University of Maryland – Baltimore, Baltimore, MD, 21201, USA

    Bryan A. Krantz

Authors
  1. Bryan A. Krantz
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bryan A. Krantz .

Editor information

Editors and Affiliations

  1. Department of Biology and Biochemistry, University of Houston, Houston, TX, USA

    Anne H. Delcour

Rights and permissions

Reprints and permissions

Copyright information

© 2015 Springer International Publishing Switzerland

About this chapter

Cite this chapter

Krantz, B.A. (2015). Anthrax Toxin Protective Antigen Forms an Unusual Channel That Unfolds and Translocates Proteins Across Membranes. In: Delcour, A.H. (eds) Electrophysiology of Unconventional Channels and Pores. Springer Series in Biophysics, vol 18. Springer, Cham. https://doi.org/10.1007/978-3-319-20149-8_9

Download citation

Publish with us

Policies and ethics

Search

Navigation