PDBsum entry 1ut2

Adhesin

PDB id: 1ut2

Contents

Protein chains

(+ 3 more) 138 a.a. *

Ligands

SO4 ×3

Waters ×27

* Residue conservation analysis

PDB id:

1ut2

Name:

Adhesin

Title:

Afae-3 adhesin from escherichia coli

Structure:

Afimbrial adhesin afa-iii. Chain: a, b, c, d, e, f, g, h, i. Synonym: afa-iii, afae3. Engineered: yes. Mutation: yes

Source:

Escherichia coli. Organism_taxid: 562. Strain: a30. Expressed in: escherichia coli. Expression_system_taxid: 562. Other_details: n-terminus 6-his-tagged

Biol. unit:

Trimer (from PDB file)

Resolution:

3.30Å R-factor: 0.228 R-free: 0.266

Authors:

K.L.Anderson,J.Billington,D.Pettigrew,E.Cota,P.Roversi,P.Simpson, H.A.Chen,P.Urvil,L.Dumerle,P.Barlow,E.Medof,R.A.G.Smith,B.Nowicki, C.Le Bouguenec,S.M.Lea,S.Matthews

Key ref:

D.Pettigrew et al. (2004). High resolution studies of the Afa/Dr adhesin DraE and its interaction with chloramphenicol. J Biol Chem, 279, 46851-46857. PubMed id: 15331605 DOI: 10.1074/jbc.M409284200

Date:

02-Dec-03 Release date: 31-Aug-04

Protein chains

Q57254  (AFAE3_ECOLX) -  Afimbrial adhesin AFA-III from Escherichia coli

Seq: 160 a.a.

Struc: 138 a.a.*

* PDB and UniProt seqs differ at 2 residue positions (black crosses)

DOI no: 10.1074/jbc.M409284200

J Biol Chem 279:46851-46857 (2004)

PubMed id: 15331605

High resolution studies of the Afa/Dr adhesin DraE and its interaction with chloramphenicol.

D.Pettigrew, K.L.Anderson, J.Billington, E.Cota, P.Simpson, P.Urvil, F.Rabuzin, P.Roversi, B.Nowicki, L.du Merle, C.Le Bouguénec, S.Matthews, S.M.Lea.

ABSTRACT

Pathogenic Escherichia coli expressing Afa/Dr adhesins are able to cause both urinary tract and diarrheal infections. The Afa/Dr adhesins confer adherence to epithelial cells via interactions with the human complement regulating protein, decay accelerating factor (DAF or CD55). Two of the Afa/Dr adhesions, AfaE-III and DraE, differ from each other by only three residues but are reported to have several different properties. One such difference is disruption of the interaction between DraE and CD55 by chloramphenicol, whereas binding of AfaE-III to CD55 is unaffected. Here we present a crystal structure of a strand-swapped trimer of wild type DraE. We also present a crystal structure of this trimer in complex with chloramphenicol, as well as NMR data supporting the binding position of chloramphenicol within the crystal. The crystal structure reveals the precise atomic basis for the sensitivity of DraE-CD55 binding to chloramphenicol and demonstrates that in contrast to other chloramphenicol-protein complexes, drug binding is mediated via recognition of the chlorine "tail" rather than via intercalation of the benzene rings into a hydrophobic pocket.

Selected figure(s)

Figure 1.
FIG. 1. Crystallographic structures for DraE and AfaE-III, comparisons to earlier structure of AfaE-dsc. a, crystal structure of AfaE-III trimer shown as a ribbon representation. Monomers are colored red, green, and blue. The G strand that is donated from another monomer to construct the native fimbrae (11) is striped, and the five residues that differ significantly in their place in the fold between the trimeric and monomerized forms of the protein are highlighted in purple. Also shown is a ribbon representation of monomers of AfaE taken either from the trimer (AfaE-III) or from the engineered monomer (AfaE-dsc) (11) oriented and colored to aid comparisons of the structures. b, overlay of worm representations of the backbones of monomers from the AfaE-III (green) and DraE (cyan) trimers. Side chains are shown for the three amino acids that differ in sequence between the two adhesins. c, topology diagrams for the adhesin trimers and for the assembled fimbrae (11). Coloring is as described in a. d, non-reducing SDS-PAGE for the fimbrial preparation of the Dr hemagglutinin (second and third lanes). Markers are shown in the first and fourth lanes and correspond to molecular weights X, Y, and Z. M, size markers; F, fimbrae.

Figure 3.
FIG. 3. Analysis of lack of chloramphenicol sensitivity of AfaE-III. a, backbone representations of the x-ray structures of DraE (cyan) and AfaE-III (green) monomers as in Fig. 1b. The -carbon positions of the three amino acids that are altered between these different strains are highlighted as numbered space filling balls. The chloramphenicol bound to the DraE is shown in a ball-and-stick representation and labeled Cm. b, analogous view of AfaE-III to that shown of DraE in Fig. 2a. The residues that differ between these adhesins are colored red. The side chain of Met-88 is seen to lie across the Cm-binding pocket. c, Cm sensitivity of CD55 binding assayed by surface plasmon resonance (see "Experimental Procedures"). Shown is CD55 (1 µM) binding to AfaE-III and DraE in the absence (-) and presence (+) of Cm (2.8 mM). Values presented are the mean and associated errors derived from three (DraE) and six (AfaE-III) repeated measurements.

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2004, 279, 46851-46857) copyright 2004.

Figures were selected by an automated process.