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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Barbey, C., Budin-Verneuil, A., Cauchard, S., Hartke, A., Laugier, C., Pichereau, V., and Petry, S. 2009. Proteomic analysis and immunogenicity of secreted proteins from Rhodococcus equi ATCC 33701. Vet. Microbiol.135, 334–345.
Bek-Thomsen, M., Tettelin, H., Hance, I., Nelson, K.E., and Kilian, M. 2008. Population diversity and dynamics of Streptococcus mitis, Streptococcus oralis, and Streptococcus infantis in the upper respiratory tracts of adults, determined by a nonculture strategy. Infect. Immun.76, 1889–1896.
Choi, C.W., Lee, Y.G., Kwon, S.O., Kim, H.Y., Lee, J.C., Chung, Y.H., Yun, C.Y., and Kim, S.I. 2012. Analysis of Streptococcus pneumoniae secreted antigens by immuno-proteomic approach. Diagn. Microbiol. Infect. Dis.72, 318–327.
Choi, C.W., Yun, S.H., Kwon, S.O., Leem, S.H., Choi, J.S., Yun, C.Y., and Kim, S.I. 2010. Analysis of cytoplasmic membrane proteome of Streptococcus pneumoniae by shotgun proteomic approach. J. Microbiol.48, 872–876.
Gomez-Gascon, L., Luque, I., Olaya-Abril, A., Jimenez-Munguia, I., Orbegozo-Medina, R.A., Peralbo, E., Tarradas, C., and Rodriguez-Ortega, M.J. 2012. Exploring the pan-surfome of Streptococcus suis: looking for common protein antigens. J. Proteomics75, 5654–5666.
Kadioglu, A., Weiser, J.N., Paton, J.C., and Andrew, P.W. 2008. The role of Streptococcus pneumoniae virulence factors in host respiratory colonization and disease. Nat. Rev. Microbiol.6, 288–301.
Lee, Y.G., Leem, S.H., Chung, Y.H., and Kim, S.I. 2012. Characterization of thermostable deblocking aminopeptidases of archaeon Thermococcus onnurineus NA1 by proteomic and biochemical approaches. J. Microbiol.50, 792–797.
Liu, Y.H., Wylie, D., Zhao, J., Cure, R., Cutler, C., Cannon-Carlson, S., Yang, X., Nagabhushan, T.L., and Pramanik, B.N. 2011. Mass spectrometric characterization of the isoforms in Escherichia coli recombinant DNA-derived interferon alpha-2b. Anal. Biochem.408, 105–117.
Mariappan, V., Vellasamy, K.M., Thimma, J.S., Hashim, O.H., and Vadivelu, J. 2010. Identification of immunogenic proteins from Burkholderia cepacia secretome using proteomic analysis. Vaccine28, 1318–1324.
Morsczeck, C., Prokhorova, T., Sigh, J., Pfeiffer, M., Bille-Nielsen, M., Petersen, J., Boysen, A., Kofoed, T., Frimodt-Moller, N., Nyborg-Nielsen, P., andet al. 2008. Streptococcus pneumoniae: proteomics of surface proteins for vaccine development. Clin. Microbiol. Infect.14, 74–81.
Olaya-Abril, A., Gomez-Gascon, L., Jimenez-Munguia, I., Obando, I., and Rodriguez-Ortega, M.J. 2012. Another turn of the screw in shaving Gram-positive bacteria: Optimization of proteomics surface protein identification in Streptococcus pneumoniae. J. Proteomics75, 3733–3746.
Pranchevicius, M.C., Oliveira, L.L., Rosa, J.C., Avanci, N.C., Quiapim, A.C., Roque-Barreira, M.C., and Goldman, M.H. 2012. Characterization and optimization of ArtinM lectin expression in Escherichia coli. BMC Biotechnol.12, 44.
Sun, X., Yang, X.Y., Yin, X.F., Yu, G., Xiao, C.L., He, X., and He, Q.Y. 2011. Proteomic analysis of membrane proteins from Streptococcus pneumoniae with multiple separation methods plus high accuracy mass spectrometry. OMICS15, 683–694.
Tettelin, H., Nelson, K.E., Paulsen, I.T., Eisen, J.A., Read, T.D., Peterson, S., Heidelberg, J., DeBoy, R.T., Haft, D.H., Dodson, R.J., andet al. 2001. Complete genome sequence of a virulent isolate of Streptococcus pneumoniae. Science293, 498–506.
Yadav, V., Panilaitis, B., Shi, H., Numuta, K., Lee, K., and Kaplan, D.L. 2011. N-acetylglucosamine 6-phosphate deacetylase (nagA) is required for N-acetyl glucosamine assimilation in Gluconacetobacter xylinus. PLoS One6, e18099.
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Choi, CW., An, HY., Lee, Y.J. et al. Characterization of Streptococcus pneumoniae N-acetylglucosamine-6-phosphate deacetylase as a novel diagnostic marker. J Microbiol. 51, 659–664 (2013). https://doi.org/10.1007/s12275-013-3451-8
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DOI: https://doi.org/10.1007/s12275-013-3451-8