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

doi:10.1107/S174430910801213X

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 journal › Article › peer-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 language	English
Pages (from-to)	512-515
Number of pages	4
Journal	Acta Crystallographica Section F: Structural Biology and Crystallization Communications
Volume	64
Issue number	6
DOIs	https://doi.org/10.1107/S174430910801213X
Publication status	Published - 2008 Jul 22
Externally published	Yes

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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Miyafusa, T., Tanaka, Y., Kuroda, M., Ohta, T., & Tsumoto, K. (2008). Expression, purification, crystallization and preliminary diffraction analysis of CapF, a capsular polysaccharide-synthesis enzyme from Staphylococcus aureus. Acta Crystallographica Section F: Structural Biology and Crystallization Communications, 64(6), 512-515. https://doi.org/10.1107/S174430910801213X

Expression, purification, crystallization and preliminary diffraction analysis of CapF, a capsular polysaccharide-synthesis enzyme from Staphylococcus aureus. / Miyafusa, Takamitsu; Tanaka, Yoshikazu; Kuroda, Makoto; Ohta, Toshiko; Tsumoto, Kouhei.

In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 64, No. 6, 22.07.2008, p. 512-515.

Research output: Contribution to journal › Article › peer-review

Miyafusa, T, Tanaka, Y, Kuroda, M, Ohta, T & Tsumoto, K 2008, 'Expression, purification, crystallization and preliminary diffraction analysis of CapF, a capsular polysaccharide-synthesis enzyme from Staphylococcus aureus', Acta Crystallographica Section F: Structural Biology and Crystallization Communications, vol. 64, no. 6, pp. 512-515. https://doi.org/10.1107/S174430910801213X

Miyafusa, Takamitsu ; Tanaka, Yoshikazu ; Kuroda, Makoto ; Ohta, Toshiko ; Tsumoto, Kouhei. / Expression, purification, crystallization and preliminary diffraction analysis of CapF, a capsular polysaccharide-synthesis enzyme from Staphylococcus aureus. In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications. 2008 ; Vol. 64, No. 6. pp. 512-515.

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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 {\AA}. The crystal belongs to space group P3221, with unit-cell parameters a = b = 119.6, c = 129.5 {\AA}.",

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