Crystallization of the F41 fragment of flagellin and data collection from extremely thin crystals

Fadel A. Samatey, Katsumi Imada, Ferenc Vonderviszt, Yasuo Shirakihara, Keiichi Namba

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Flagellin, which constructs supercoiled filaments of the bacterial flagellum, is very difficult to crystallize because of its strong tendency to polymerize. We therefore crystallized the F41 fragment of flagellin, which does not polymerize because terminal regions that play important roles in polymerization are cleaved off. F41 was crystallized by the hanging drop vapor diffusion method in a mixture of polyethylene glycol, glycerol, and isopropanol, with a reservoir solution covered with silicon oil. The two key factors for success in growing sufficiently large crystals were isopropanol and silicon oil, which worked well to reduce the otherwise very high nucleation rate that resulted in hundreds of tiny crystals. The crystals were grown to very thin plates with thickness less than 10 μm, which made the collection of diffraction data very difficult. Freezing and annealing of the crystals and irradiation at synchrotron beamlines had to be carried out by specific methods and under specific conditions for its structure analysis at 2.0-Å resolution.

Original languageEnglish
Pages (from-to)106-111
Number of pages6
JournalJournal of Structural Biology
Volume132
Issue number2
DOIs
Publication statusPublished - Jan 1 2000

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Keywords

  • Bacterial flagellum
  • Cryocrystallography
  • Crystallization
  • Flagellin fragment
  • Salmonella
  • Thin crystal
  • X-ray data collection

ASJC Scopus subject areas

  • Structural Biology

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