Open AccessExpression, purification, crystallization and preliminary diffraction analysis of CapF, a capsular polysaccharide‐synthesis enzyme from Staphylococcus aureus
Author(s) -
Miyafusa Takamitsu,
Tanaka Yoshikazu,
Kuroda Makoto,
Ohta Toshiko,
Tsumoto Kouhei
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910801213x
Subject(s) - crystallization , enzyme , staphylococcus aureus , transferase , polysaccharide , biochemistry , chemistry , biology , microbiology and biotechnology , bacteria , organic chemistry , genetics
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 P 3 2 21, with unit‐cell parameters a = b = 119.6, c = 129.5 Å.