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Methylation, crystallization and SAD phasing of the Csu pilus CsuC–CsuE chaperone–adhesin subunit pre‐assembly complex from Acinetobacter baumannii
Author(s) -
Pakharukova Natalia,
Tuittila Minna,
Paavilainen Sari,
Zavialov Anton
Publication year - 2017
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x17009566
Subject(s) - pilus , chaperone (clinical) , fimbriae proteins , protein subunit , bacterial adhesin , microbiology and biotechnology , crystallography , biofilm , acinetobacter baumannii , periplasmic space , chemistry , escherichia coli , biophysics , bacteria , biology , biochemistry , genetics , medicine , pathology , pseudomonas aeruginosa , gene
Acinetobacter baumannii is one of the most difficult Gram‐negative bacteria to control and treat. This pathogen forms biofilms on hospital surfaces and medical devices using Csu pili assembled via the archaic chaperone–usher pathway. To uncover the mechanism of bacterial attachment to abiotic surfaces, it was aimed to determine the crystal structure of the pilus tip adhesin CsuE. The CsuC–CsuE chaperone–subunit pre‐assembly complex was purified from the periplasm of Escherichia coli overexpressing CsuC and CsuE. Despite the high purity of the complex, no crystals could be obtained. This challenge was solved by the methylation of lysine residues. The complex was crystallized in 0.1 M bis‐tris pH 5.5, 17% PEG 3350 using the hanging‐drop vapour‐diffusion method. The crystals diffracted to a resolution of 2.31 Å and belonged to the triclinic space group P 1, with unit‐cell parameters a = 53.84, b = 63.85, c = 89.25 Å, α = 74.65, β = 79.65, γ = 69.07°. Initial phases were derived from a single anomalous diffraction experiment using a selenomethionine derivative.