PDBsum entry 2omy

Cell invasion/cell adhesion

PDB id: 2omy

Contents

Protein chains

461 a.a. *

105 a.a. *

Metals

_CL ×4

_CA ×2

Waters ×736

* Residue conservation analysis

PDB id:

2omy

Name:

Cell invasion/cell adhesion

Title:

Crystal structure of inla s192n/hec1 complex

Structure:

Internalin-a. Chain: a. Fragment: internalin domain. Engineered: yes. Mutation: yes. Epithelial-cadherin. Chain: b. Fragment: n-terminal domain of human e-cadherin. Engineered: yes

Source:

Listeria monocytogenes. Organism_taxid: 169963. Strain: egd-e. Gene: inla. Expressed in: escherichia coli bl21. Expression_system_taxid: 511693. Homo sapiens. Human. Organism_taxid: 9606.

Resolution:

1.70Å R-factor: 0.159 R-free: 0.211

Authors:

T.Wollert,D.W.Heinz,W.D.Schubert

Key ref:

T.Wollert et al. (2007). Extending the host range of Listeria monocytogenes by rational protein design. Cell, 129, 891-902. PubMed id: 17540170 DOI: 10.1016/j.cell.2007.03.049

Date:

23-Jan-07 Release date: 05-Jun-07

Protein chain

P0DJM0  (INLA_LISMO) -  Internalin A from Listeria monocytogenes serovar 1/2a (strain ATCC BAA-679 / EGD-e)

Seq: 800 a.a.

Struc: 461 a.a.*

Protein chain

P12830  (CADH1_HUMAN) -  Cadherin-1 from Homo sapiens

Seq: 882 a.a.

Struc: 105 a.a.*

* PDB and UniProt seqs differ at 6 residue positions (black crosses)

DOI no: 10.1016/j.cell.2007.03.049

Cell 129:891-902 (2007)

PubMed id: 17540170

Extending the host range of Listeria monocytogenes by rational protein design.

T.Wollert, B.Pasche, M.Rochon, S.Deppenmeier, J.van den Heuvel, A.D.Gruber, D.W.Heinz, A.Lengeling, W.D.Schubert.

ABSTRACT

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.

Selected figure(s)

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].

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.

The above figures are reprinted by permission from Cell Press: Cell (2007, 129, 891-902) copyright 2007.

Figures were selected by the author.

Author's comment

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 human diseases therefore most often involve existing pathogens whose arsenal has been altered to allow them to infect previously inaccessible hosts. We have simulated this chance occurrence by extending the host range of the human pathogen Listeria monocytogenes via the intestine to include the mouse. Amino acid substitutions in the listerial invasion protein InlA that increase the affinity for its receptor human E-cadherin were identified based on the recognition complex between these two proteins and verified by isothermal titration calorimetry. 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, a versatile murine model of human listeriosis has thus been created.