title: 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.
abstract: 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 C222(1), 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.
authors: Department of Chemistry, University of Turku, Joint Biotechnology Laboratory, BioCity, Tykistökatu 6A, 20520 Turku, Finland.
authors: Natalia
authors: N
authors: Pakharukova
authors: Minna
authors: M
authors: Tuittila
authors: Anton
authors: A
authors: Zavialov
mesh_terms: D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D003460:Crystallization; D018360:Crystallography, X-Ray; D004196:Disease Outbreaks; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D036781:Fimbriae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005858:Germany / Q000453:epidemiology; D006463:Hemolytic-Uremic Syndrome / Q000453:epidemiology / Q000382:microbiology; D006801:Humans; D008969:Molecular Sequence Data; D021122:Protein Subunits / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D054323:Shiga-Toxigenic Escherichia coli / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D013455:Sulfur / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: C079725:AggA protein, E coli; D029968:Escherichia coli Proteins; D021122:Protein Subunits; D011994:Recombinant Proteins; D036781:Fimbriae Proteins; D013455:Sulfur
keywords: H4; Shiga toxin-producing Escherichia coli O104; aggregative adherence fimbriae; chaperone/usher pathway; sulfur single anomalous diffraction