Acta Biomedica Scientifica (Sep 2016)

Detection of membrane-bound proteases of Francisella tularensis

  • A. V. Korneva,
  • V. B. Nikolaev,
  • C. N. Kozlov,
  • E. Y. Markov,
  • A. V. Mazepa,
  • Y. O. Popova

DOI
https://doi.org/10.12737/23416
Journal volume & issue
Vol. 1, no. 5
pp. 155 – 159

Abstract

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Sterile preparations of membrane fractions were prepared by processing of live Francisella tularensis cells of different subspecies with 4.5 M urea solution and differential centrifugation. For the first time, proteolytic activity was detected and studied by tests of radial enzyme diffusion and substrate polyacrylamide gel electrophoresis using gelatin as a substrate. Spectrum of gelatinases in the resulted preparation were detected. Quantitative inter-strain differences in the protease activities and their qualitative composition in membrane preparations of various virulent F. tularensis strains was analyzed. Avirulent F. tularensis 21/400 subsp. holarctica (1-214) strain demonstrated the greatest gelatinase activity in enzyme diffusion method and the lowest hydrolytic activity was seen in F. tularensis B-399 A-Cole subsp. tularensis (1-386) and F. tularensis Utah 112 subsp. novicida (1-384), other preparations showed intermediate activity. Enzyme electrophoresis in the protease spectra determined the presence of proteins with proteases activity 50-100 kDa, and in the spectrum preparations of F. tularensis 1-386 and 1-384 were detected additional bands of proteases.

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