IDENTIFICATION OF IMMUNOGENIC PROTEINS OF STRAINS OF BACILLUS ANTHRACIS IN MALDI TOF MS

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Abstract

Aim. Identification of obtained in host-simulated conditions immunogenic proteins of isogenic variants of Bacillus anthracis 575/122. Materials and methods. We used culture filtrate of isogenic variants of B. anthracis 575/122: R02 (pXOL pXO2+); R01 (pXO1+ pXO2‘); R00 (pXOL pX02~), obtained in host-simulated conditions. In the one-dimensional polyacrylamide gel electrophoresis and immunoblotting with hyperimmune serums immunodominant proteins, that have been identified in MALDI TOF MS. Results. Immunoblotting revealed proteins with molecular masses in range 97 - 14.1 kDa. 90 kDa protein from strain B. anthracis 575/122 R01 in MALDI TOF MS was identified as protective antigen with 85.810 kDa. Protein with molecular mass 60 kDa was identified as GMP synthase with molecular mass 57.239 kDa. In the culture filtrates of three strains two common antigen were identified: protein with molecular mass 97 kDa, identified as B. anthracis EA 1 with molecular mass 91.361 kDa protein and 45 kDa protein as enolase B. anthracis with molecular mass 46.418 kDa. Conclusion. Thus, the conditions that simulate the host can promote the production of immunodominant proteins of B. anthracis. The data about molecular-weight characteristics of protective antigen and EA 1 protein as well as some of proteases of B. anthracis are confirmed by the MALDI TOF MS. The results can be used for isolation of these proteins to improve the diagnostic and vaccine preparations.

About the authors

M. P. Chervakova

Volgograd State Research Institute for Plague Control

Author for correspondence.
Russian Federation

T. N. Sharov

Volgograd State Research Institute for Plague Control

Russian Federation

LA. .. Barkova

Volgograd State Research Institute for Plague Control

Russian Federation

A. M. Barkov

Volgograd State Research Institute for Plague Control

Russian Federation

D. V. Viktorov

Volgograd State Research Institute for Plague Control

Russian Federation

A. V. Toporkov

Volgograd State Research Institute for Plague Control

Russian Federation

References

  1. Барков А.М., Новоженина А.В., Порохня С.В., Ткаченко Г.А., Липницкий А.В. Характеристика изогенных вариантов Bacillus anthracis с различным содержанием плазмид вирулентности. Журн. микробиол. 2015, 1:17-22.
  2. Баркова И.А., Червакова М.П., Барков А.М., Новоженина А.В., Викторов Д.В. Диагностическое значение некоторых иммунодоминантных белков изогенных вариантов Bacillus anthracis. Клиническая лабораторная диагностика. 2016, 61 (12): 833838.
  3. Краснов Н.В., Лютвинский Я.И., Подольская Е.П. Масс-спектрометрия с мягкими методами ионизации в протеомном анализе. Научное приборостроение. 2010, 20 (4): 5-20.
  4. Семенов Б.Ф., Зверев В.В., Хаитов Р.М. Прогноз развития вакцинопрофилактики в первые десятилетия XXI века. Иммунология. 2009,6: 324-335.
  5. Agarwal S., Kulshreshtha R, Bambah Mukku D. et al. Alpha-Enolase binds to human plasminogen on the surface of Bacillus anthracis. Biochim. Biophys. Acta. 2008,1784 (7-8): 986994.
  6. Boyer A. E., Gallegos-Candela M., Lins R.C. et al. Quantitative mass spectrometry for bacterial protein toxins - a sensitive, specific, high-throughput tool for detection and diagnosis. Molecules. 2011, 16: 2391-2413.
  7. Chitlaru T., Gat O., Grosfeld H. et al. Identification ofin vivo-expressed immunogenic proteins by serological proteome analysis of the bacillus anthracis secretome. Infect. Immun. 2007,75: 2841-2852.
  8. Chitlaru X, Zaide G., Ehrlich S. et al. HtrA is a major virulence determinant of Bacillus anthracis. Molecular Microbiology. 2011, 81 (6): 1542-1559.
  9. Fouet A. AtxA, a Bacillus anthracis global virulence regulator. Res. Microbiol. 2010, 161: 735-742.
  10. LamonicaJ.M., Wagner M., EschenbrennerM. etal. Comparative secretome analyses ofthree Bacillus anthracis strains with variant plasmid contents. Infect. Immun. 2005, 73: 36463658.
  11. Liu X., Wang D., Ren J. et al. Identification of the immunogenic spore and vegetative proteins of Bacillus anthracis vaccine strain A16R. PLoS ONE. 2013, 8 (3): e57959.
  12. Pflughoeft K.J., Swick M.C., Engler D. A. et al. Modulation of the Bacillus anthracis secretome by the immune inhibitor Al protease. J. Bacteriol. 2014,196 (2): 424-435.
  13. Ristroph J.D., Ivins B.E. Elaboration of Bacillus anthracis antigens in a new, defined culture medium. Infect. Immun. 1983, 39:483-486.
  14. Shevchenko A., Tomas H., Havlis J. et al. In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nature protocols. 2006, 1 (6): 2856-2860.
  15. Walz A., Mujer C., Connolly J. et al. Bacillus anthracis secretome time course under host-simulated conditions and identification of immunogenic proteins. Proteome Science. 2007,5:

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Copyright (c) 2018 Chervakova M.P., Sharov T.N., Barkova L..., Barkov A.M., Viktorov D.V., Toporkov A.V.

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