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Acta Crystallogr F Struct Biol Commun. 2017 Mar 1;73(Pt 3):123-129. doi: 10.1107/S2053230X17002096. Epub 2017 Feb 21.

Crystallization of FcpA from Leptospira, a novel flagellar protein that is essential for pathogenesis.

Author information

1
Laboratory of Molecular and Structural Microbiology, Institut Pasteur de Montevideo, 11400 Montevideo, Uruguay.
2
Department of Epidemiology of Microbial Disease, Yale School of Public Health, New Haven, CT 06520, USA.
3
Unit of Biology of Spirochetes, Department of Microbiology, Institut Pasteur, 25-28 Rue du Dr Roux, 75015 Paris, France.

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.

KEYWORDS:

FcpA; Leptospira biflexa; Leptospira interrogans; SAD phasing; flagella; leptospirosis; motility; spirochetes

PMID:
28291747
PMCID:
PMC5349305
DOI:
10.1107/S2053230X17002096
[Indexed for MEDLINE]
Free PMC Article

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