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Contents |
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(+ 8 more)
552 a.a.
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(+ 8 more)
181 a.a.
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* Residue conservation analysis
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PDB id:
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Toxin receptor/toxin
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Title:
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Crystal structure of the anthrax toxin protective antigen heptameric prepore bound to the vwa domain of cmg2, an anthrax toxin receptor
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Structure:
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Protective antigen. Chain: a, b, c, d, e, f, g, h, i, j, k, l, m, o. Fragment: 63-kda domain. Synonym: pxo1-110. Pa-83. Pa83. Engineered: yes. Anthrax toxin receptor 2. Chain: a, b, c, d, e, f, g, h, i, j, k, l, m, o. Fragment: vwa domain. Synonym: capillary morphogenesis protein 2. Cmg-2.
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Source:
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Bacillus anthracis str.. Organism_taxid: 191218. Strain: a2012. Gene: paga. Expressed in: escherichia coli. Expression_system_taxid: 562. Homo sapiens. Human. Organism_taxid: 9606.
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Biol. unit:
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Not given
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Resolution:
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4.30Å
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R-factor:
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0.322
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R-free:
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0.330
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Authors:
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D.B.Lacy,D.J.Wigelsworth,R.A.Melnyk,R.J.Collier
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Key ref:
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D.B.Lacy
et al.
(2004).
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.
PubMed id:
DOI:
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Date:
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10-Jul-04
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Release date:
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17-Aug-04
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PROCHECK
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Headers
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References
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DOI no:
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Proc Natl Acad Sci U S A
101:13147-13151
(2004)
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PubMed id:
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Structure of heptameric protective antigen bound to an anthrax toxin receptor: a role for receptor in pH-dependent pore formation.
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D.B.Lacy,
D.J.Wigelsworth,
R.A.Melnyk,
S.C.Harrison,
R.J.Collier.
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ABSTRACT
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After binding to cellular receptors and proteolytic activation, the protective
antigen component of anthrax toxin forms a heptameric prepore. The prepore later
undergoes pH-dependent conversion to a pore, mediating translocation of the
edema and lethal factors to the cytosol. We describe structures of the prepore
(3.6 A) and a prepore:receptor complex (4.3 A) that reveal the location of
pore-forming loops and an unexpected interaction of the receptor with the
pore-forming domain. Lower pH is required for prepore-to-pore conversion in the
presence of the receptor, indicating that this interaction regulates
pH-dependent pore formation. We present an example of a receptor negatively
regulating pH-dependent membrane insertion.
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Selected figure(s)
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Figure 1.
Fig. 1. The (PA[63])[7] prepore structure. (a) A single
monomer from the 3.6-Å (PA[63])[7] structure. Domains 1,
2, 3, and 4 are colored in pink, green, yellow and blue,
respectively. The previously unresolved 303-322 (red), 343-350
(blue) and 512-515 (blue) loops are visible whereas the 275-283
and 426-427 loops remain unstructured (black dotted lines). (b)
An aerial view (domain 1' is at the top, closest to the viewer)
of the PA[63] heptamer with one monomer colored as in a. Domain
2 lines the prepore lumen whereas domains 3 and 4 are located on
the outside of the heptamer ring. (c) Domains 2 (green) and 4
(blue) from the 3.6-Å (PA[63])[7] structure, as viewed
from the bottom. The domain 2 insertion loop (red) projects out
to bind the neighboring monomer in a groove between domains 2
and 4. (d) The domain 2 insertion loop contacts domain 4 from
its own monomer (residues 600-602) and domains 2 (residue 414)
and 4 (residues 668-670) from the neighboring monomer. N306 was
mutated to cysteine and labeled with pyrene for the experiments
described in Fig. 3b. Residues F313 and F314 are predicted to
form the tip of the membrane inserted  -hairpin (10).
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Figure 2.
Fig. 2. The (PA[63])[7](CMG2)[7] structure. (a) In these
bottom and side views, the CMG2 VWA domains are depicted in
pink. Only three PA:CMG2 monomers are shown in the side view for
clarity. (b) The CMG2 VWA domain (pink) binds both PA domain 2
(white, green, and dark blue) and PA domain 4 (light and dark
blue). Direct contacts within the interface are depicted in dark
pink (CMG2) and dark blue (PA). The PA insertion loop and the
contiguous 2 2 and 2 3 -strands
(green) are predicted to peel away from the domain 2 -barrel
core to form a pore. The CMG2 VWA domain bound to the PA 340-348
loop is likely to impede this rearrangement. (c) This close-up
of the PA:CMG2 interface is colored as in b. Previous PA
mutagenesis studies suggest the importance of residues G342,
W346, I656, N682, and D683 (blue) in intoxication, although the
G342C and W346C mutants may reflect structural instability (16).
The domain 2 R344 (blue) is buried within the PA:CMG2 interface
and may form a salt bridge with CMG2 E122 (pink). Residues Y119,
H121, E122, and Y158 of CMG2 (pink) are strictly conserved in
ATR/TEM8 and cluster at the PA domain 2 interface, suggesting
that ATR/TEM8 binding will also block PA insertion loop
rearrangement. Other CMG2 residues at the interface with PA
(yellow) are not conserved in ATR/TEM8.
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Figures were
selected
by an automated process.
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Literature references that cite this PDB file's key reference
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PubMed id
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Reference
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D.Basilio,
L.D.Jennings-Antipov,
K.S.Jakes,
and
A.Finkelstein
(2011).
Trapping a translocating protein within the anthrax toxin channel: implications for the secondary structure of permeating proteins.
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J Gen Physiol,
137,
343-356.
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K.L.Thoren,
and
B.A.Krantz
(2011).
The unfolding story of anthrax toxin translocation.
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Mol Microbiol,
80,
588-595.
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L.D.Jennings-Antipov,
L.Song,
and
R.J.Collier
(2011).
Interactions of anthrax lethal factor with protective antigen defined by site-directed spin labeling.
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Proc Natl Acad Sci U S A,
108,
1868-1873.
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S.R.Crowe,
L.Garman,
R.J.Engler,
A.D.Farris,
J.D.Ballard,
J.B.Harley,
and
J.A.James
(2011).
Anthrax vaccination induced anti-lethal factor IgG: fine specificity and neutralizing capacity.
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Vaccine,
29,
3670-3678.
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A.F.Kintzer,
H.J.Sterling,
I.I.Tang,
E.R.Williams,
and
B.A.Krantz
(2010).
Anthrax toxin receptor drives protective antigen oligomerization and stabilizes the heptameric and octameric oligomer by a similar mechanism.
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PLoS One,
5,
e13888.
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B.E.Janowiak,
A.Fischer,
and
R.J.Collier
(2010).
Effects of introducing a single charged residue into the phenylalanine clamp of multimeric anthrax protective antigen.
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J Biol Chem,
285,
8130-8137.
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G.K.Feld,
K.L.Thoren,
A.F.Kintzer,
H.J.Sterling,
I.I.Tang,
S.G.Greenberg,
E.R.Williams,
and
B.A.Krantz
(2010).
Structural basis for the unfolding of anthrax lethal factor by protective antigen oligomers.
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Nat Struct Mol Biol,
17,
1383-1390.
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PDB code:
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J.Sun,
and
R.J.Collier
(2010).
Disulfide bonds in the ectodomain of anthrax toxin receptor 2 are required for the receptor-bound protective-antigen pore to function.
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PLoS One,
5,
e10553.
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S.Fu,
X.Tong,
C.Cai,
Y.Zhao,
Y.Wu,
Y.Li,
J.Xu,
X.C.Zhang,
L.Xu,
W.Chen,
and
Z.Rao
(2010).
The structure of tumor endothelial marker 8 (TEM8) extracellular domain and implications for its receptor function for recognizing anthrax toxin.
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PLoS One,
5,
e11203.
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PDB code:
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A.F.Kintzer,
K.L.Thoren,
H.J.Sterling,
K.C.Dong,
G.K.Feld,
I.I.Tang,
T.T.Zhang,
E.R.Williams,
J.M.Berger,
and
B.A.Krantz
(2009).
The protective antigen component of anthrax toxin forms functional octameric complexes.
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J Mol Biol,
392,
614-629.
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PDB code:
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A.S.Williams,
S.Lovell,
A.Anbanandam,
R.El-Chami,
and
J.G.Bann
(2009).
Domain 4 of the anthrax protective antigen maintains structure and binding to the host receptor CMG2 at low pH.
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Protein Sci,
18,
2277-2286.
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PDB code:
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G.Vernier,
J.Wang,
L.D.Jennings,
J.Sun,
A.Fischer,
L.Song,
and
R.J.Collier
(2009).
Solubilization and characterization of the anthrax toxin pore in detergent micelles.
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Protein Sci,
18,
1882-1895.
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G.van der Goot,
and
J.A.Young
(2009).
Receptors of anthrax toxin and cell entry.
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Mol Aspects Med,
30,
406-412.
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J.N.Tournier,
R.G.Ulrich,
A.Quesnel-Hellmann,
M.Mohamadzadeh,
and
B.G.Stiles
(2009).
Anthrax, toxins and vaccines: a 125-year journey targeting Bacillus anthracis.
|
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Expert Rev Anti Infect Ther,
7,
219-236.
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J.Oscherwitz,
F.Yu,
J.L.Jacobs,
T.H.Liu,
P.R.Johnson,
and
K.B.Cease
(2009).
Synthetic peptide vaccine targeting a cryptic neutralizing epitope in domain 2 of Bacillus anthracis protective antigen.
|
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Infect Immun,
77,
3380-3388.
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M.Rajapaksha,
J.F.Eichler,
J.Hajduch,
D.E.Anderson,
K.L.Kirk,
and
J.G.Bann
(2009).
Monitoring anthrax toxin receptor dissociation from the protective antigen by NMR.
|
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Protein Sci,
18,
17-23.
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M.Y.Go,
E.M.Chow,
and
J.Mogridge
(2009).
The cytoplasmic domain of anthrax toxin receptor 1 affects binding of the protective antigen.
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Infect Immun,
77,
52-59.
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|
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R.J.Collier
(2009).
Membrane translocation by anthrax toxin.
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Mol Aspects Med,
30,
413-422.
|
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S.Sharma,
D.Thomas,
J.Marlett,
M.Manchester,
and
J.A.Young
(2009).
Efficient neutralization of antibody-resistant forms of anthrax toxin by a soluble receptor decoy inhibitor.
|
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Antimicrob Agents Chemother,
53,
1210-1212.
|
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|
|
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A.Fischer,
D.J.Mushrush,
D.B.Lacy,
and
M.Montal
(2008).
Botulinum neurotoxin devoid of receptor binding domain translocates active protease.
|
| |
PLoS Pathog,
4,
e1000245.
|
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A.Sivasubramanian,
J.A.Maynard,
and
J.J.Gray
(2008).
Modeling the structure of mAb 14B7 bound to the anthrax protective antigen.
|
| |
Proteins,
70,
218-230.
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|
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D.S.Anderson,
and
R.O.Blaustein
(2008).
Preventing voltage-dependent gating of anthrax toxin channels using engineered disulfides.
|
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J Gen Physiol,
132,
351-360.
|
|
|
|
|
|
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H.Katayama,
B.E.Janowiak,
M.Brzozowski,
J.Juryck,
S.Falke,
E.P.Gogol,
R.J.Collier,
and
M.T.Fisher
(2008).
GroEL as a molecular scaffold for structural analysis of the anthrax toxin pore.
|
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Nat Struct Mol Biol,
15,
754-760.
|
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|
|
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J.G.Smedley,
J.S.Sharp,
J.F.Kuhn,
and
K.B.Tomer
(2008).
Probing the pH-dependent prepore to pore transition of Bacillus anthracis protective antigen with differential oxidative protein footprinting.
|
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Biochemistry,
47,
10694-10704.
|
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J.Sun,
A.E.Lang,
K.Aktories,
and
R.J.Collier
(2008).
Phenylalanine-427 of anthrax protective antigen functions in both pore formation and protein translocation.
|
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Proc Natl Acad Sci U S A,
105,
4346-4351.
|
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|
|
|
|
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M.Vuyisich,
S.Gnanakaran,
J.A.Lovchik,
C.R.Lyons,
and
G.Gupta
(2008).
A dual-purpose protein ligand for effective therapy and sensitive diagnosis of anthrax.
|
| |
Protein J,
27,
292-302.
|
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|
|
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|
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M.Yan,
M.H.Roehrl,
E.Basar,
and
J.Y.Wang
(2008).
Selection and evaluation of the immunogenicity of protective antigen mutants as anthrax vaccine candidates.
|
| |
Vaccine,
26,
947-955.
|
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|
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N.Abboud,
and
A.Casadevall
(2008).
Immunogenicity of Bacillus anthracis protective antigen domains and efficacy of elicited antibody responses depend on host genetic background.
|
| |
Clin Vaccine Immunol,
15,
1115-1123.
|
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P.Roblin,
V.Guillet,
O.Joubert,
D.Keller,
M.Erard,
L.Maveyraud,
G.Prévost,
and
L.Mourey
(2008).
A covalent S-F heterodimer of leucotoxin reveals molecular plasticity of beta-barrel pore-forming toxins.
|
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Proteins,
71,
485-496.
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PDB code:
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S.C.Taft,
and
A.A.Weiss
(2008).
Toxicity of anthrax toxin is influenced by receptor expression.
|
| |
Clin Vaccine Immunol,
15,
1330-1336.
|
|
|
|
|
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A.Fokine,
V.D.Bowman,
A.J.Battisti,
Q.Li,
P.R.Chipman,
V.B.Rao,
and
M.G.Rossmann
(2007).
Cryo-electron microscopy study of bacteriophage T4 displaying anthrax toxin proteins.
|
| |
Virology,
367,
422-427.
|
|
|
|
|
|
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B.S.Powell,
J.T.Enama,
W.J.Ribot,
W.Webster,
S.Little,
T.Hoover,
J.J.Adamovicz,
and
G.P.Andrews
(2007).
Multiple asparagine deamidation of Bacillus anthracis protective antigen causes charge isoforms whose complexity correlates with reduced biological activity.
|
| |
Proteins,
68,
458-479.
|
|
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|
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|
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D.J.Manayani,
D.Thomas,
K.A.Dryden,
V.Reddy,
M.E.Siladi,
J.M.Marlett,
G.J.Rainey,
M.E.Pique,
H.M.Scobie,
M.Yeager,
J.A.Young,
M.Manchester,
and
A.Schneemann
(2007).
A viral nanoparticle with dual function as an anthrax antitoxin and vaccine.
|
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PLoS Pathog,
3,
1422-1431.
|
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|
|
|
|
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H.M.Scobie,
J.M.Marlett,
G.J.Rainey,
D.B.Lacy,
R.J.Collier,
and
J.A.Young
(2007).
Anthrax toxin receptor 2 determinants that dictate the pH threshold of toxin pore formation.
|
| |
PLoS ONE,
2,
e329.
|
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|
|
|
|
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J.A.Young,
and
R.J.Collier
(2007).
Anthrax toxin: receptor binding, internalization, pore formation, and translocation.
|
| |
Annu Rev Biochem,
76,
243-265.
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K.H.Chen,
S.Liu,
L.A.Bankston,
R.C.Liddington,
and
S.H.Leppla
(2007).
Selection of anthrax toxin protective antigen variants that discriminate between the cellular receptors TEM8 and CMG2 and achieve targeting of tumor cells.
|
| |
J Biol Chem,
282,
9834-9845.
|
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|
|
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|
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S.Liu,
H.J.Leung,
and
S.H.Leppla
(2007).
Characterization of the interaction between anthrax toxin and its cellular receptors.
|
| |
Cell Microbiol,
9,
977-987.
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B.DeLaBarre,
and
A.T.Brunger
(2006).
Considerations for the refinement of low-resolution crystal structures.
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| |
Acta Crystallogr D Biol Crystallogr,
62,
923-932.
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F.Tama,
G.Ren,
C.L.Brooks,
and
A.K.Mitra
(2006).
Model of the toxic complex of anthrax: responsive conformational changes in both the lethal factor and the protective antigen heptamer.
|
| |
Protein Sci,
15,
2190-2200.
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|
|
H.M.Scobie,
D.J.Wigelsworth,
J.M.Marlett,
D.Thomas,
G.J.Rainey,
D.B.Lacy,
M.Manchester,
R.J.Collier,
and
J.A.Young
(2006).
Anthrax toxin receptor 2-dependent lethal toxin killing in vivo.
|
| |
PLoS Pathog,
2,
e111.
|
|
|
|
|
|
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H.M.Scobie,
and
J.A.Young
(2006).
Divalent metal ion coordination by residue T118 of anthrax toxin receptor 2 is not essential for protective antigen binding.
|
| |
PLoS ONE,
1,
e99.
|
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|
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|
|
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I.I.Salles,
D.E.Voth,
S.C.Ward,
K.M.Averette,
R.K.Tweten,
K.A.Bradley,
and
J.D.Ballard
(2006).
Cytotoxic activity of Bacillus anthracis protective antigen observed in a macrophage cell line overexpressing ANTXR1.
|
| |
Cell Microbiol,
8,
1272-1281.
|
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|
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|
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M.Gao,
and
K.Schulten
(2006).
Onset of anthrax toxin pore formation.
|
| |
Biophys J,
90,
3267-3279.
|
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|
M.J.Gubbins,
J.D.Berry,
C.R.Corbett,
J.Mogridge,
X.Y.Yuan,
L.Schmidt,
B.Nicolas,
A.Kabani,
and
R.S.Tsang
(2006).
Production and characterization of neutralizing monoclonal antibodies that recognize an epitope in domain 2 of Bacillus anthracis protective antigen.
|
| |
FEMS Immunol Med Microbiol,
47,
436-443.
|
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|
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|
|
|
M.L.Peterson,
and
P.M.Schlievert
(2006).
Glycerol monolaurate inhibits the effects of Gram-positive select agents on eukaryotic cells.
|
| |
Biochemistry,
45,
2387-2397.
|
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|
|
|
|
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R.A.Melnyk,
and
R.J.Collier
(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.
|
|
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|
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S.J.Tilley,
and
H.R.Saibil
(2006).
The mechanism of pore formation by bacterial toxins.
|
| |
Curr Opin Struct Biol,
16,
230-236.
|
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|
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|
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B.Chen,
E.M.Vogan,
H.Gong,
J.J.Skehel,
D.C.Wiley,
and
S.C.Harrison
(2005).
Determining the structure of an unliganded and fully glycosylated SIV gp120 envelope glycoprotein.
|
| |
Structure,
13,
197-211.
|
|
|
|
|
|
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D.B.Lacy,
H.C.Lin,
R.A.Melnyk,
O.Schueler-Furman,
L.Reither,
K.Cunningham,
D.Baker,
and
R.J.Collier
(2005).
A model of anthrax toxin lethal factor bound to protective antigen.
|
| |
Proc Natl Acad Sci U S A,
102,
16409-16414.
|
|
|
|
|
|
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G.J.Rainey,
D.J.Wigelsworth,
P.L.Ryan,
H.M.Scobie,
R.J.Collier,
and
J.A.Young
(2005).
Receptor-specific requirements for anthrax toxin delivery into cells.
|
| |
Proc Natl Acad Sci U S A,
102,
13278-13283.
|
|
|
|
|
|
|
H.M.Scobie,
and
J.A.Young
(2005).
Interactions between anthrax toxin receptors and protective antigen.
|
| |
Curr Opin Microbiol,
8,
106-112.
|
|
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J.Y.Wang,
and
M.H.Roehrl
(2005).
Anthrax vaccine design: strategies to achieve comprehensive protection against spore, bacillus, and toxin.
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Med Immunol,
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L.Abrami,
N.Reig,
and
F.G.van der Goot
(2005).
Anthrax toxin: the long and winding road that leads to the kill.
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Trends Microbiol,
13,
72-78.
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R.J.Gilbert
(2005).
Inactivation and activity of cholesterol-dependent cytolysins: what structural studies tell us.
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| |
Structure,
13,
1097-1106.
|
|
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R.Mabry,
M.Rani,
R.Geiger,
G.B.Hubbard,
R.Carrion,
K.Brasky,
J.L.Patterson,
G.Georgiou,
and
B.L.Iverson
(2005).
Passive protection against anthrax by using a high-affinity antitoxin antibody fragment lacking an Fc region.
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Infect Immun,
73,
8362-8368.
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R.Ramachandran,
R.K.Tweten,
and
A.E.Johnson
(2005).
The domains of a cholesterol-dependent cytolysin undergo a major FRET-detected rearrangement during pore formation.
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| |
Proc Natl Acad Sci U S A,
102,
7139-7144.
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S.Zhang,
A.Finkelstein,
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(2004).
Evidence that translocation of anthrax toxin's lethal factor is initiated by entry of its N terminus into the protective antigen channel.
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Proc Natl Acad Sci U S A,
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The most recent references are shown first.
Citation data come partly from CiteXplore and partly
from an automated harvesting procedure. Note that this is likely to be
only a partial list as not all journals are covered by
either method. However, we are continually building up the citation data
so more and more references will be included with time.
Where a reference describes a PDB structure, the PDB
code is
shown on the right.
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Links
