Expression, purification, crystallization and preliminary diffraction analysis of CapF, a capsular polysaccharide-synthesis enzyme from Staphylococcus aureus

Takamitsu Miyafusa, Yoshikazu Tanaka, Makoto Kuroda, Toshiko Ohta, Kouhei Tsumoto

Research output: Contribution to journalArticlepeer-review

4 Citations (Scopus)

Abstract

Capsular polysaccharides (CPs) are important virulence factors of Staphylococcus aureus. The biosynthesis of type 5 and type 8 CPs (CP5 and CP8), which are produced by most clinical isolates of S. aureus, is catalyzed by 16 CP-assembling proteins. One of these proteins is the enzyme CapF, which catalyzes the synthesis of UDP-N-acetyl-l-fucosamine, a component of both CP5 and CP8. Here, the cloning, expression, purification, crystallization and diffraction analysis of CapF are reported. Optimization of the crystallization conditions by differential scanning calorimetry afforded a crystal of selenomethionine-substituted CapF that diffracted to a resolution of 2.80 Å. The crystal belongs to space group P3221, with unit-cell parameters a = b = 119.6, c = 129.5 Å.

Original languageEnglish
Pages (from-to)512-515
Number of pages4
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume64
Issue number6
DOIs
Publication statusPublished - 2008 Jul 22
Externally publishedYes

Keywords

  • Capsular polysaccharide-synthesis enzymes
  • Differential scanning calorimetry
  • Staphylococcus aureus

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

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