Crystallization and sulfur SAD phasing of AggA, the major subunit of aggregative adherence fimbriae type I from the Escherichia coli strain that caused an outbreak of haemolytic-uraemic syndrome in Germany

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Vol. 69; no. 12; pp. 1389 - 1392
• Main Authors: Pakharukova, Natalia, Tuittila, Minna, Zavialov, Anton
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01-12-2013; Wiley Subscription Services, Inc
• Subjects: aggregative adherence fimbriae; Amino Acid Sequence; Bacterial Adhesion; chaperone/usher pathway; Crystallization; Crystallization Communications; Crystallography, X-Ray; Disease Outbreaks; Escherichia coli; Escherichia coli Proteins - chemistry; Escherichia coli Proteins - genetics; Escherichia coli Proteins - metabolism; Fimbriae Proteins - chemistry; Fimbriae Proteins - genetics; Fimbriae Proteins - metabolism; Gene Expression; Germany - epidemiology; Hemolytic-Uremic Syndrome - epidemiology; Hemolytic-Uremic Syndrome - microbiology; Humans; Molecular Sequence Data; Protein Subunits - chemistry; Protein Subunits - genetics; Protein Subunits - metabolism; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Shiga toxin-producing Escherichia coli O104; Shiga-Toxigenic Escherichia coli - chemistry; Shiga-Toxigenic Escherichia coli - genetics; Shiga-Toxigenic Escherichia coli - metabolism; Shiga-Toxigenic Escherichia coli - pathogenicity; Sulfur - chemistry; sulfur single anomalous diffraction; X-Ray Diffraction; Germany; Shiga toxin-producing Escherichia coli O104; chaperone/usher pathway; H4; sulfur single anomalous diffraction; aggregative adherence fimbriae
• ISSN: 1744-3091; 1744-3091; 2053-230X
• DOI: 10.1107/S1744309113029990

The outbreak of Shiga toxin‐producing Escherichia coli O104:H4 infection in Germany in 2011 was associated with significant mortality and morbidity owing to the progressive development of haemolytic‐uraemic syndrome. The outbreak strain emerged recently as a result of horizontal transfer events leading to the acquisition of a number of virulence factors. Among them, aggregative adherence fimbriae type I (AAF/I) are considered to be particularly important since they are involved in the initial attachment of bacteria to the intestinal mucosa. Here, the crystallization and preliminary X‐ray diffraction analysis of the major subunit of AAF/I, AggA, are reported. Crystallization of recombinant donor‐strand complemented AggA was performed by the vapour‐diffusion method. The crystals diffracted to 1.55 Å resolution and belonged to the orthorhombic space group C2221, with unit‐cell parameters a = 77.83, b = 80.17, c = 91.42 Å. Despite a low sulfur content of the protein [0.57%(w/w)], sufficiently accurate initial phases were derived from a sulfur SAD experiment.