Open AccessStructure of the catalytic domain of Streptococcus pneumoniae sialidase NanA
Author(s) -
Xu Guogang,
Li Xuejun,
Andrew Peter W.,
Taylor Garry L.
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309108024044
Subject(s) - sialidase , glycoconjugate , enzyme , neuraminic acid , sialic acid , biochemistry , virulence , yeast , streptococcus pneumoniae , biology , chemistry , microbiology and biotechnology , stereochemistry , gene , neuraminidase , antibiotics
Streptococcus pneumoniae genomes encode three sialidases, NanA, NanB and NanC, which are key virulence factors that remove sialic acids from various glycoconjugates. The enzymes have potential as drug targets and also as vaccine candidates. The 115 kDa NanA is the largest of the three sialidases and is anchored to the bacterial membrane. Although recombinantly expressed full‐length NanA was soluble, it failed to crystallize; therefore, a 56.5 kDa domain that retained full enzyme activity was subcloned. The purified enzyme was crystallized in 0.1 M MES pH 6.5, 30%( w / v ) PEG 4000 using the sitting‐drop vapour‐diffusion method. Data were collected at 100 K to 2.5 Å resolution from a crystal grown in the presence of the inhibitor 2‐deoxy‐2,3‐dehydro‐ N ‐acetyl neuraminic acid. The crystal belongs to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 49.2, b = 95.6, c = 226.6 Å. The structure was solved by molecular replacement and refined to final R and R free factors of 0.246 and 0.298, respectively.