Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis

Bourdeau, Raymond W; Malito, Enrico; Chenal, Alexandre; Bishop, Brian L; Musch, Mark W; Villereal, Mitch L; Chang, Eugene B; Mosser, Elise M; Rest, Richard F; Tang, Wei-Jen

doi:10.1074/jbc.M807631200

Title: Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis

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Abstract

Anthrolysin O (ALO) is a pore-forming, cholesterol-dependent cytolysin (CDC) secreted by Bacillus anthracis, the etiologic agent for anthrax. Growing evidence suggests the involvement of ALO in anthrax pathogenesis. Here, we show that the apical application of ALO decreases the barrier function of human polarized epithelial cells as well as increases intracellular calcium and the internalization of the tight junction protein occludin. Using pharmacological agents, we also found that barrier function disruption requires increased intracellular calcium and protein degradation. We also report a crystal structure of the soluble state of ALO. Based on our analytical ultracentrifugation and light scattering studies, ALO exists as a monomer. Our ALO structure provides the molecular basis as to how ALO is locked in a monomeric state, in contrast to other CDCs that undergo antiparallel dimerization or higher order oligomerization in solution. ALO has four domains and is globally similar to perfringolysin O (PFO) and intermedilysin (ILY), yet the highly conserved undecapeptide region in domain 4 (D4) adopts a completely different conformation in all three CDCs. Consistent with the differences within D4 and at the D2-D4 interface, we found that ALO D4 plays a key role in affecting the barrier function of C2BBE cells, whereas PFOmore »« less

Authors:: Bourdeau, Raymond W; Malito, Enrico; Chenal, Alexandre; Bishop, Brian L; Musch, Mark W; Villereal, Mitch L; Chang, Eugene B; Mosser, Elise M; Rest, Richard F; Tang, Wei-Jen

Publication Date:: Tue Jun 02 00:00:00 EDT 2009

Research Org.:: Argonne National Lab. (ANL), Argonne, IL (United States)

Sponsoring Org.:: USDOE

OSTI Identifier:: 1005645

Resource Type:: Journal Article

Journal Name:: J. Biol. Chem.

Additional Journal Information:: Journal Volume: 284; Journal Issue: (21) ; 05, 2009; Journal ID: ISSN 0021-9258

Country of Publication:: United States

Language:: ENGLISH

Subject:: 36 MATERIALS SCIENCE; BACILLUS; CALCIUM; CRYSTAL STRUCTURE; DIMERIZATION; ELEMENTS; FUNCTIONS; HUMAN POPULATIONS; LIGHT SCATTERING; PATHOGENESIS; PROTEINS; ULTRACENTRIFUGATION

Citation Formats


                    Bourdeau, Raymond W, Malito, Enrico, Chenal, Alexandre, Bishop, Brian L, Musch, Mark W, Villereal, Mitch L, Chang, Eugene B, Mosser, Elise M, Rest, Richard F, Tang, Wei-Jen, CNRS-UMR), Drexel-MED), and UC). Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis.  United States: N. p., 2009. 
        Web.  doi:10.1074/jbc.M807631200.

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                    Bourdeau, Raymond W, Malito, Enrico, Chenal, Alexandre, Bishop, Brian L, Musch, Mark W, Villereal, Mitch L, Chang, Eugene B, Mosser, Elise M, Rest, Richard F, Tang, Wei-Jen, CNRS-UMR), Drexel-MED), & UC). Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis.  United States.  https://doi.org/10.1074/jbc.M807631200

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                    Bourdeau, Raymond W, Malito, Enrico, Chenal, Alexandre, Bishop, Brian L, Musch, Mark W, Villereal, Mitch L, Chang, Eugene B, Mosser, Elise M, Rest, Richard F, Tang, Wei-Jen, CNRS-UMR), Drexel-MED), and UC). 2009.  
        "Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis".  United States.  https://doi.org/10.1074/jbc.M807631200.

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@article{osti_1005645,

  title        = {Cellular Functions and X-ray Structure of Anthrolysin O, a Cholesterol-dependent Cytolysin Secreted by Bacillus anthracis},

  author       = {Bourdeau, Raymond W and Malito, Enrico and Chenal, Alexandre and Bishop, Brian L and Musch, Mark W and Villereal, Mitch L and Chang, Eugene B and Mosser, Elise M and Rest, Richard F and Tang, Wei-Jen and CNRS-UMR) and Drexel-MED) and UC)},

  abstractNote = {Anthrolysin O (ALO) is a pore-forming, cholesterol-dependent cytolysin (CDC) secreted by Bacillus anthracis, the etiologic agent for anthrax. Growing evidence suggests the involvement of ALO in anthrax pathogenesis. Here, we show that the apical application of ALO decreases the barrier function of human polarized epithelial cells as well as increases intracellular calcium and the internalization of the tight junction protein occludin. Using pharmacological agents, we also found that barrier function disruption requires increased intracellular calcium and protein degradation. We also report a crystal structure of the soluble state of ALO. Based on our analytical ultracentrifugation and light scattering studies, ALO exists as a monomer. Our ALO structure provides the molecular basis as to how ALO is locked in a monomeric state, in contrast to other CDCs that undergo antiparallel dimerization or higher order oligomerization in solution. ALO has four domains and is globally similar to perfringolysin O (PFO) and intermedilysin (ILY), yet the highly conserved undecapeptide region in domain 4 (D4) adopts a completely different conformation in all three CDCs. Consistent with the differences within D4 and at the D2-D4 interface, we found that ALO D4 plays a key role in affecting the barrier function of C2BBE cells, whereas PFO domain 4 cannot substitute for this role. Novel structural elements and unique cellular functions of ALO revealed by our studies provide new insight into the molecular basis for the diverse nature of the CDC family.},

  doi          = {10.1074/jbc.M807631200},

  url          = {https://www.osti.gov/biblio/1005645},
  journal      = {J. Biol. Chem.},

  issn         = {0021-9258},

  number       = (21) ; 05, 2009,

  volume       = 284,

  place        = {United States},

  year         = {Tue Jun 02 00:00:00 EDT 2009},

  month        = {Tue Jun 02 00:00:00 EDT 2009}

}

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