title: Crystallization of FcpA from Leptospira, a novel flagellar protein that is essential for pathogenesis.
abstract: The protein FcpA is a unique component of the flagellar filament of spirochete bacteria belonging to the genus Leptospira. Although it plays an essential role in translational motility and pathogenicity, no structures of FcpA homologues are currently available in the PDB. Its three-dimensional structure will unveil the novel motility mechanisms that render pathogenic Leptospira particularly efficient at invading and disseminating within their hosts, causing leptospirosis in humans and animals. FcpA from L. interrogans was purified and crystallized, but despite laborious attempts no useful X ray diffraction data could be obtained. This challenge was solved by expressing a close orthologue from the related saprophytic species L. biflexa. Three different crystal forms were obtained: a primitive and a centred monoclinic form, as well as a hexagonal variant. All forms diffracted X-rays to suitable resolutions for crystallographic analyses, with the hexagonal type typically reaching the highest limits of 2.0 Å and better. A variation of the quick-soaking procedure resulted in an iodide derivative that was instrumental for single-wavelength anomalous diffraction methods.
authors.affiliation: Laboratory of Molecular and Structural Microbiology, Institut Pasteur de Montevideo, 11400 Montevideo, Uruguay.
authors.forename: Fabiana
authors.initials: F
authors.lastname: San Martin
authors.affiliation: Laboratory of Molecular and Structural Microbiology, Institut Pasteur de Montevideo, 11400 Montevideo, Uruguay.
authors.forename: Ariel E
authors.initials: AE
authors.lastname: Mechaly
authors.affiliation: Laboratory of Molecular and Structural Microbiology, Institut Pasteur de Montevideo, 11400 Montevideo, Uruguay.
authors.forename: Nicole
authors.initials: N
authors.lastname: Larrieux
authors.affiliation: Department of Epidemiology of Microbial Disease, Yale School of Public Health, New Haven, CT 06520, USA.
authors.forename: Elsio A
authors.initials: EA
authors.lastname: Wunder
authors.affiliation: Department of Epidemiology of Microbial Disease, Yale School of Public Health, New Haven, CT 06520, USA.
authors.forename: Albert I
authors.initials: AI
authors.lastname: Ko
authors.affiliation: Unit of Biology of Spirochetes, Department of Microbiology, Institut Pasteur, 25-28 Rue du Dr Roux, 75015 Paris, France.
authors.forename: Mathieu
authors.initials: M
authors.lastname: Picardeau
authors.affiliation: Laboratory of Molecular and Structural Microbiology, Institut Pasteur de Montevideo, 11400 Montevideo, Uruguay.
authors.forename: Felipe
authors.initials: F
authors.lastname: Trajtenberg
authors.affiliation: Laboratory of Molecular and Structural Microbiology, Institut Pasteur de Montevideo, 11400 Montevideo, Uruguay.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Buschiazzo
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005407:Flagella / Q000737:chemistry* / Q000378:metabolism; D015870:Gene Expression; D007919:Leptospira / Q000737:chemistry* / Q000378:metabolism; D007921:Leptospira interrogans / Q000737:chemistry* / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins
keywords: FcpA; Leptospira biflexa; Leptospira interrogans; SAD phasing; flagella; leptospirosis; motility; spirochetes