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Characterization of Streptococcus pneumoniae N-acetylglucosamine-6-phosphate deacetylase as a novel diagnostic marker
Authors:Chi-Won Choi  Hee-Young An  Yong Ju Lee  Yeol Gyun Lee  Sung Ho Yun  Edmond Changkyun Park  Yeonhee Hong  Gun-Hwa Kim  Jae-Eun Park  Sun Jong Baek  Hyun Sik Kim  Seung Il Kim
Institution:1. Division of Life Science, Korea Basic Science Institute (KBSI), Daejeon 305-806, Republic of Korea
2. Division of Mass Spectrometry Research, Korea Basic Science Institute, Ochang, 363-883, Republic of Korea
3. Department of Functional Genomics, Universtiy of Science and Technology (UST), Daejeon, 305-350, Republic of Korea
Abstract:The identification of novel diagnostic markers of pathogenic bacteria is essential for improving the accuracy of diagnoses and for developing targeted vaccines. Streptococcus pneumoniae is a significant human pathogenic bacterium that causes pneumonia. N-acetylglucosamine-6-phosphate deacetylase (NagA) was identified in a protein mixture secreted by S. pneumoniae and its strong immunogenicity was confirmed in an immuno-proteomic assay against the anti-serum of the secreted protein mixture. In this study, recombinant S. pneumoniae NagA protein was expressed and purified to analyze its protein characteristics, immunospecificity, and immunogenicity, thereby facilitating its evaluation as a novel diagnostic marker for S. pneumoniae. Mass spectrometry analysis showed that S. pneumoniae NagA contains four internal disulfide bonds and that it does not undergo post-translational modification. S. pneumoniae NagA antibodies successfully detected NagA from different S. pneumoniae strains, whereas NagA from other pathogenic bacteria species was not detected. In addition, mice infected with S. pneumoniae generated NagA antibodies in an effective manner. These results suggest that NagA has potential as a novel diagnostic marker for S. pneumoniae because of its high immunogenicity and immunospecificity.
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