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

Autor: Alejandro Buschiazzo, Ariel E. Mechaly, Nicole Larrieux, Albert I. Ko, Fabiana San Martin, Elsio A. Wunder, Mathieu Picardeau, Felipe Trajtenberg
Přispěvatelé: Molecular and structural microbiology / Microbiología Molecular y Estructural [Montevideo], Institut Pasteur de Montevideo, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Yale School of Public Health (YSPH), Fundação Oswaldo Cruz (FIOCRUZ), Réseau International des Instituts Pasteur (RIIP), Biologie des Spirochètes / Biology of Spirochetes, Institut Pasteur [Paris], Integrative Microbiology of Zoonotic Agents [Paris and Montevideo] (IMiZA), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut Pasteur [Paris], Fundação Oswaldo Cruz / Oswaldo Cruz Foundation (FIOCRUZ), Institut Pasteur [Paris] (IP), Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut Pasteur [Paris] (IP)
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
MESH: Leptospira interrogans
MESH: Sequence Homology
Amino Acid
Protein Data Bank (RCSB PDB)
Gene Expression
MESH: Amino Acid Sequence
Crystallography
X-Ray
medicine.disease_cause
Biochemistry
MESH: Leptospira
MESH: Recombinant Proteins
Plasmid
Structural Biology
leptospirosis
spirochetes
Cloning
Molecular
Peptide sequence
MESH: Bacterial Proteins
Leptospira
biology
Chemistry
MESH: Escherichia coli
Condensed Matter Physics
Recombinant Proteins
motility
SAD phasing
flagella
Leptospira interrogans
Plasmids
MESH: Gene Expression
FcpA
Biophysics
MESH: Sequence Alignment
Sequence alignment
Flagellum
Leptospira biflexa
MESH: Flagella
Article
03 medical and health sciences
Bacterial Proteins
MESH: Plasmids
Escherichia coli
Genetics
medicine
[CHIM.CRIS]Chemical Sciences/Cristallography
MESH: Cloning
Molecular
Amino Acid Sequence
[SDV.BBM.BC]Life Sciences [q-bio]/Biochemistry
Molecular Biology/Biochemistry [q-bio.BM]
Sequence Homology
Amino Acid
biology.organism_classification
MESH: Crystallography
X-Ray
[SDV.MP.BAC]Life Sciences [q-bio]/Microbiology and Parasitology/Bacteriology
030104 developmental biology
Sequence Alignment
Zdroj: Acta crystallographica. Section F, Structural biology communications
Acta crystallographica. Section F, Structural biology communications, John Wiley & Sons Ltd, 2017, 73 (3), pp.123-129. ⟨10.1107/S2053230X17002096⟩
Acta crystallographica Section F : Structural biology communications [2014-...]
Acta crystallographica Section F : Structural biology communications [2014-..], 2017, 73 (3), pp.123-129. ⟨10.1107/S2053230X17002096⟩
ISSN: 2053-230X
Popis: The protein FcpA is a unique component of the flagellar filament of spirochete bacteria belonging to the genusLeptospira. 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 pathogenicLeptospiraparticularly efficient at invading and disseminating within their hosts, causing leptospirosis in humans and animals. FcpA fromL. interroganswas 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 speciesL. 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.
Databáze: OpenAIRE
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