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PDBsum entry 2omv
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Cell invasion/cell adhesion
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PDB id
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2omv
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References listed in PDB file
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Key reference
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Title
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Extending the host range of listeria monocytogenes by rational protein design.
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Authors
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T.Wollert,
B.Pasche,
M.Rochon,
S.Deppenmeier,
J.Van den heuvel,
A.D.Gruber,
D.W.Heinz,
A.Lengeling,
W.D.Schubert.
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Ref.
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Cell, 2007,
129,
891-902.
[DOI no: ]
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PubMed id
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Abstract
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In causing disease, pathogens outmaneuver host defenses through a dedicated
arsenal of virulence determinants that specifically bind or modify individual
host molecules. This dedication limits the intruder to a defined range of hosts.
Newly emerging diseases mostly involve existing pathogens whose arsenal has been
altered to allow them to infect previously inaccessible hosts. We have emulated
this chance occurrence by extending the host range accessible to the human
pathogen Listeria monocytogenes by the intestinal route to include the mouse.
Analyzing the recognition complex of the listerial invasion protein InlA and its
human receptor E-cadherin, we postulated and verified amino acid substitutions
in InlA to increase its affinity for E-cadherin. Two single substitutions
increase binding affinity by four orders of magnitude and extend binding
specificity to include formerly incompatible murine E-cadherin. By rationally
adapting a single protein, we thus create a versatile murine model of human
listeriosis.
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Figure 1.
Figure 1. Re-engineered Variants of Internalin (InlA) in
Complex with the N-Terminal Domain of Human E-Cadherin
(A)
Superposition of InlA/hEC1 (violet) and InlA^S192N-Y369S/hEC1
(gray).
(B) Critical residues near mutation site
S192N[InlA] are shown as ball and stick. Asn192[InlA] adds a
direct H bond to the carbonyl oxygen of Phe17[hEC1].
(C)
The mutation Y369S[InlA] introduces a water-bridged interaction
to Asn27[hEC1].
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Figure 5.
Figure 5. Survival of Mice Intragastrically Infected with
Lmo-EGD and Lmo-InlA^m and Associated Bacterial Organ Loads over
Time
Survival curves (A) and organ loads (B–F) of female
C57BL/6J mice inoculated intragastrically with Lmo-EGD (dashed
curve, ○) or Lmo-InlA^m (solid curves, ●). (A) shows that
Lmo-InlA^m exhibit more than 1000-fold higher virulence than
wild-type Lmo-EGD (inocula as indicated, n = 10 for each
bacterial strain and experiment). (B–F) shows that 1 ×
10^10 bacteria of either strain were administered
intragastrically to analyze kinetics of bacterial growth (n = 6
for 24 h p.i. and n = 12 for all others). Organ loads were
ascertained at five time points in the stomach (B), small
intestine (C), mesenteric lymph nodes (D), spleen (E), and liver
(F). All data are from two independent experiments. The bar
represents the median for each time point and genotype; 95%
confidence intervals are indicated. Statistical significance by
Mann-Whitney U nonparametric test: ^*p < 0.05, ^**p < 0.01, and
^***p < 0.001.
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The above figures are
reprinted
by permission from Cell Press:
Cell
(2007,
129,
891-902)
copyright 2007.
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