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Stress proteins in the cytoplasmic membrane fraction of Bacillus subtilis

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Abstract

Stress proteomes of the cytoplasmic membrane fraction of Bacillus subtilis trp C2-exposed to acid pH and ethanol were characterized. Although these stress factors impair the cell function in a specific manner, they share the ability to denature proteins. Therefore, specific and general stress proteins in the membranes were investigated. Both ethanol (3 %) and pH 5.0 increase the doubling time from 17 to 25 min. Isolated cytoplasmic membranes were subjected to an optimized 2D PAGE analysis which permitted the separation and analysis of ≈450 distinct protein spots. Two alternative methods of protein detection were compared, i.e. silver staining and 35S-l-methionine pulse labeling; the stress induced proteins were identified by MALDI-TOF MS. After ethanol stress, five proteins were increased, viz. YdaP, Ctc, YfhM, YjcH and YwaC. Acid stress proteins were AcoB, YkwC, SodA, YjcH and YwaC. Proteins YjcH and YwaC were increased after ethanol as well as acid pH treatment.

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Abbreviations

CHAPS:

3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate

DTT:

1,4-dithiothreitol

IPG:

immobilized pH gradient

MALDI:

matrix-assisted laser desorption/ionization

M :

molar mass

MS:

mass spectrometry

PAGE:

polyacrylamide gel electrophoresis

PMSF:

phenylmethanesulfonyl fluoride

SDS:

sodium dodecyl sulfate

TOF:

time-of-flight

µ:

growth rate

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Authors and Affiliations

  1. Department of Genetics and Microbiology, Faculty of Science, Charles University in Prague, 120 00, Prague, Czech Republic

    D. Petráčková, L. Šemberová & J. Svobodová

  2. Institute of Microbiology v.v.i., Academy of Sciences of the Czech Republic, Prague, Czech Republic

    D. Petráčková & P. Halada

  3. Institute of Physiology v.v.i., Academy of Sciences of the Czech Republic, Prague, Czech Republic

    P. Svoboda

  4. Institute of Microbiology ASCR v.v.i., 142 20, Prague 4, Czech Republic

    D. Petráčková

Authors
  1. D. Petráčková
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  2. L. Šemberová
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  3. P. Halada
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  4. P. Svoboda
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  5. J. Svobodová
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Correspondence to D. Petráčková.

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Petráčková, D., Šemberová, L., Halada, P. et al. Stress proteins in the cytoplasmic membrane fraction of Bacillus subtilis . Folia Microbiol 55, 427–434 (2010). https://doi.org/10.1007/s12223-010-0072-z

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  • DOI: https://doi.org/10.1007/s12223-010-0072-z

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