TY - JOUR
T1 - Expression, purification, crystallization and preliminary diffraction analysis of CapF, a capsular polysaccharide-synthesis enzyme from Staphylococcus aureus
AU - Miyafusa, Takamitsu
AU - Tanaka, Yoshikazu
AU - Kuroda, Makoto
AU - Ohta, Toshiko
AU - Tsumoto, Kouhei
PY - 2008
Y1 - 2008
N2 - 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.
AB - 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.
KW - Capsular polysaccharide-synthesis enzymes
KW - Differential scanning calorimetry
KW - Staphylococcus aureus
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U2 - 10.1107/S174430910801213X
DO - 10.1107/S174430910801213X
M3 - Article
C2 - 18540063
AN - SCOPUS:46249092301
SN - 1744-3091
VL - 64
SP - 512
EP - 515
JO - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
JF - Acta Crystallographica Section F: Structural Biology and Crystallization Communications
IS - 6
ER -