Development of a Method for the Extraction of the Total Proteome of Bacillus anthracis Spores

Authors

  • E.A. Koteneva Stavropol Research Anti-Plague Institute, 13-15, Sovetskaya str., Stavropol, 355035 Russia; North Caucasus Federal University", Stavropol, 355009 Russia
  • O.I. Tsygankova Stavropol Research Anti-Plague Institute, 13-15, Sovetskaya str., Stavropol, 355035 Russia
  • A.V. Kalinin Stavropol Research Anti-Plague Institute, 13-15, Sovetskaya str., Stavropol, 355035 Russia
  • V.Yu. Shcherbakova Stavropol Research Anti-Plague Institute, 13-15, Sovetskaya str., Stavropol, 355035 Russia
  • I.S. Rodionov Stavropol Research Anti-Plague Institute, 13-15, Sovetskaya str., Stavropol, 355035 Russia
  • V.V. Serdyukov Stavropol Research Anti-Plague Institute, 13-15, Sovetskaya str., Stavropol, 355035 Russia
  • A.V. Abramovich Stavropol Research Anti-Plague Institute, 13-15, Sovetskaya str., Stavropol, 355035 Russia

DOI:

https://doi.org/10.18097/BMCRM00154

Keywords:

2D-gel electrophoresis; disintegration; lysis solutions; extraction of the proteomic complex; spore proteome; Bacillus anthracis

Abstract

We have developed a method for obtaining the total proteome of Bacillus anthracis spores, which combines efficient protein extraction with reliable disinfection of samples. We used 7 strains of B. anthracis: 4 plasmid-free, 3 diplasmid, one of which with an atypical type of capsule formation in air. The schemes for isolating the total spore proteome were tested using various lysis solutions in the presence of bacterial protease inhibitors, with the inclusion of the stages of treatment of spores with trichloroacetic acid and mechanical disintegration and with their exclusion. The quality and completeness of the extraction of the total proteome of the spores of the samples was assessed in one-dimensional (1D) and twodimensional (2D) electrophoresis. Treatment of spores with trichloroacetic acid increased the reliability of material disinfection and reduces the loss of the final product. Mechanical disintegration after treatment of spores with lysing solutions increases the completeness of extraction of spore proteins in a wide range of molecular weights and facilitated the process of sterilizing filtration. Final filtration of the lysate through a PVDF filter with a pore size of 0.22 μm provided additional decontamination of samples without reducing their quality. Thus, the use of the proposed sample preparation scheme makes it possible to obtain complete protein extracts of spores of B. anthracis strains, suitable for comparative analysis of the proteome and search for a possible correlation with the features phenotypic properties and mechanisms that ensure the preservation of the pathogen anthrax in environmental objects, including soil

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Published

2021-12-23

How to Cite

Koteneva, E., Tsygankova, O., Kalinin, A., Shcherbakova, V., Rodionov, I., Serdyukov, V., & Abramovich, A. (2021). Development of a Method for the Extraction of the Total Proteome of Bacillus anthracis Spores. Biomedical Chemistry: Research and Methods, 4(4), e00154. https://doi.org/10.18097/BMCRM00154

Issue

Section

EXPERIMENTAL RESEARCH