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Hormone/growth factor
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PDB id
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1rj7
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* Residue conservation analysis
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PDB id:
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Hormone/growth factor
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Title:
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Crystal structure of eda-a1
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Structure:
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Ectodysplasin a. Chain: a, b, d, e, f, g, h, i, j, k, l, m. Fragment: tnf domain of eda-a1. Synonym: ectodermal dysplasia protein, eda protein. Engineered: yes
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Source:
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Homo sapiens. Human. Organism_taxid: 9606. Gene: ed1, eda. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.
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Biol. unit:
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Trimer (from
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Resolution:
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2.30Å
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R-factor:
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0.201
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R-free:
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0.260
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Authors:
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S.G.Hymowitz,D.M.Compaan,M.Yan,H.Ackerly,V.M.Dixit,M.A.Staro A.M.De Vos
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Key ref:
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M.Mourez
et al.
(2003).
Mapping dominant-negative mutations of anthrax protective antigen by scanning mutagenesis.
Proc Natl Acad Sci U S A,
100,
13803-13808.
PubMed id:
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Date:
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18-Nov-03
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Release date:
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09-Dec-03
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PROCHECK
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Headers
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References
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Q92838
(EDA_HUMAN) -
Ectodysplasin-A
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Seq:
Struc:
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391 a.a.
144 a.a.
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Key: |
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PfamA domain |
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PfamB domain |
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Secondary structure |
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CATH domain |
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Gene Ontology (GO) functional annotation
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Cellular component
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membrane
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1 term
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Biological process
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immune response
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1 term
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Biochemical function
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protein binding
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2 terms
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Proc Natl Acad Sci U S A
100:13803-13808
(2003)
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PubMed id:
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Mapping dominant-negative mutations of anthrax protective antigen by scanning mutagenesis.
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M.Mourez,
M.Yan,
D.B.Lacy,
L.Dillon,
L.Bentsen,
A.Marpoe,
C.Maurin,
E.Hotze,
D.Wigelsworth,
R.A.Pimental,
J.D.Ballard,
R.J.Collier,
R.K.Tweten.
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ABSTRACT
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The protective antigen (PA) moiety of anthrax toxin transports edema factor and
lethal factor to the cytosol of mammalian cells by a mechanism that depends on
its ability to oligomerize and form pores in the endosomal membrane. Previously,
some mutated forms of PA, designated dominant negative (DN), were found to
coassemble with wild-type PA and generate defective heptameric pore-precursors
(prepores). Prepores containing DN-PA are impaired in pore formation and in
translocating edema factor and lethal factor across the endosomal membrane. To
create a more comprehensive map of sites within PA where a single amino acid
replacement can give a DN phenotype, we used automated systems to generate a
Cys-replacement mutation for each of the 568 residues of PA63, the active 63-kDa
proteolytic fragment of PA. Thirty-three mutations that reduced PA's ability to
mediate toxicity at least 100-fold were identified in all four domains of PA63.
A majority (22) were in domain 2, the pore-forming domain. Seven of the domain-2
mutations, located in or adjacent to the 2beta6 strand, the 2beta7 strand, and
the 2beta10-2beta11 loop, gave the DN phenotype. This study demonstrates the
feasibility of high-throughput scanning mutagenesis of a moderate sized protein.
The results show that DN mutations cluster in a single domain and implicate
2beta6 and 2beta7 strands and the 2beta10-2beta11 loop in the conformational
rearrangement of the prepore to the pore. They also add to the repertoire of
mutations available for structure-function studies and for designing new
antitoxic agents for treatment of anthrax.
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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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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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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.Wu,
C.Feng,
Y.Hong,
A.Guo,
S.Cao,
J.Dong,
L.Lin,
and
Z.Liu
(2010).
Soluble expression and purification of the anthrax protective antigen in E. coli and identification of a novel dominant-negative mutant N435C.
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Appl Microbiol Biotechnol, 87,
609-616.
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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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B.E.Janowiak,
A.Finkelstein,
and
R.J.Collier
(2009).
An approach to characterizing single-subunit mutations in multimeric prepores and pores of anthrax protective antigen.
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Protein Sci, 18,
348-358.
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F.Vahedi,
M.Sankian,
and
M.Mahmoudi
(2009).
Functional expression of Bacillus anthracis protective antigen in E. coli.
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Appl Biochem Biotechnol, 157,
554-561.
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S.Cao,
A.Guo,
Z.Liu,
Y.Tan,
G.Wu,
C.Zhang,
Y.Zhao,
and
H.Chen
(2009).
Investigation of new dominant-negative inhibitors of anthrax protective antigen mutants for use in therapy and vaccination.
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Infect Immun, 77,
4679-4687.
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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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T.M.Pelish,
and
M.S.McClain
(2009).
Dominant-negative inhibitors of the Clostridium perfringens epsilon-toxin.
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J Biol Chem, 284,
29446-29453.
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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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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.
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Vaccine, 26,
947-955.
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J.A.Young,
and
R.J.Collier
(2007).
Anthrax toxin: receptor binding, internalization, pore formation, and translocation.
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Annu Rev Biochem, 76,
243-265.
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|
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J.A.Melton-Witt,
L.M.Bentsen,
and
R.K.Tweten
(2006).
Identification of functional domains of Clostridium septicum alpha toxin.
|
| |
Biochemistry, 45,
14347-14354.
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|
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K.A.Christensen,
B.A.Krantz,
and
R.J.Collier
(2006).
Assembly and disassembly kinetics of anthrax toxin complexes.
|
| |
Biochemistry, 45,
2380-2386.
|
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|
|
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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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|
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V.J.Torres,
M.S.McClain,
and
T.L.Cover
(2006).
Mapping of a domain required for protein-protein interactions and inhibitory activity of a Helicobacter pylori dominant-negative VacA mutant protein.
|
| |
Infect Immun, 74,
2093-2101.
|
|
|
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|
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B.A.Aulinger,
M.H.Roehrl,
J.J.Mekalanos,
R.J.Collier,
and
J.Y.Wang
(2005).
Combining anthrax vaccine and therapy: a dominant-negative inhibitor of anthrax toxin is also a potent and safe immunogen for vaccines.
|
| |
Infect Immun, 73,
3408-3414.
|
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|
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D.A.Rasko,
M.R.Altherr,
C.S.Han,
and
J.Ravel
(2005).
Genomics of the Bacillus cereus group of organisms.
|
| |
FEMS Microbiol Rev, 29,
303-329.
|
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|
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M.Qa'dan,
K.A.Christensen,
L.Zhang,
T.M.Roberts,
and
R.J.Collier
(2005).
Membrane insertion by anthrax protective antigen in cultured cells.
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| |
Mol Cell Biol, 25,
5492-5498.
|
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|
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|
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A.R.Hoffmaster,
J.Ravel,
D.A.Rasko,
G.D.Chapman,
M.D.Chute,
C.K.Marston,
B.K.De,
C.T.Sacchi,
C.Fitzgerald,
L.W.Mayer,
M.C.Maiden,
F.G.Priest,
M.Barker,
L.Jiang,
R.Z.Cer,
J.Rilstone,
S.N.Peterson,
R.S.Weyant,
D.R.Galloway,
T.D.Read,
T.Popovic,
and
C.M.Fraser
(2004).
Identification of anthrax toxin genes in a Bacillus cereus associated with an illness resembling inhalation anthrax.
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| |
Proc Natl Acad Sci U S A, 101,
8449-8454.
|
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|
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D.B.Lacy,
D.J.Wigelsworth,
R.A.Melnyk,
S.C.Harrison,
and
R.J.Collier
(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.
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PDB codes:
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|
|
E.Santelli,
L.A.Bankston,
S.H.Leppla,
and
R.C.Liddington
(2004).
Crystal structure of a complex between anthrax toxin and its host cell receptor.
|
| |
Nature, 430,
905-908.
|
|
|
PDB code:
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|
|
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|
|
G.J.Rainey,
and
J.A.Young
(2004).
Antitoxins: novel strategies to target agents of bioterrorism.
|
| |
Nat Rev Microbiol, 2,
721-726.
|
|
|
|
|
|
|
H.Barth,
K.Aktories,
M.R.Popoff,
and
B.G.Stiles
(2004).
Binary bacterial toxins: biochemistry, biology, and applications of common Clostridium and Bacillus proteins.
|
| |
Microbiol Mol Biol Rev, 68,
373.
|
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|
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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
codes are
shown on the right.
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Links
