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A two-dimensional protein map of Pleurotus ostreatus microsomes-proteome dynamics

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Abstract

Recent studies documented that several processes in filamentous fungi are connected with microsomal enzyme activities. In this work, microsomal subproteomes of Pleurotus ostreatus were analyzed by two-dimensional (2-D) polyacrylamide gel electrophoresis and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry analysis. To assess proteome dynamics, microsomal proteins were isolated from fungal cultures after 7 and 12 days of cultivation. Additionally, 10 mg/L of 17α-ethinylestradiol (EE2) was treated with the cultures during 2 days. Despite the EE2 degradation by the fungus reached 97 and 76.3 % in 7- and 12-day-old cultures, respectively, only a minor effect on the composition of microsomal proteins was observed. The changes in protein maps related to ageing prevailed over those induced by EE2. Epoxide hydrolase, known to metabolize EE2, was detected in 12-day-old cultures only which suggests differences in EE2 degradation pathways utilized by fungal cultures of different age. The majority (32 %) of identified microsomal proteins were parts of mitochondrial energy metabolism.

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Acknowledgments

This work was supported by the project A/13/07824 of the German Academic Exchange Service, the grant no. 15-02328S of the Czech Science Foundation, and the Institutional Research Concept RVO: 61388971. In this work, instrumental equipment provided by C4Sys infrastructure was used.

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Authors and Affiliations

  1. Laboratory of Cell Signalization, Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Prague, Czech Republic

    Denisa Petráčková & Silvia Bezoušková

  2. Laboratory of Molecular Structure Characterization, Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Prague, Czech Republic

    Petr Halada

  3. Laboratory of Environmental Biotechnology, Institute of Microbiology, v.v.i., Academy of Sciences of the Czech Republic, Vídeňská 1083, 14220, Prague, Czech Republic

    Zdena Křesinová & Kateřina Svobodová

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  1. Denisa Petráčková
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  2. Petr Halada
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  3. Silvia Bezoušková
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  4. Zdena Křesinová
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  5. Kateřina Svobodová
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Corresponding author

Correspondence to Kateřina Svobodová.

Electronic supplementary material

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Fig. S1

Raw images of 2-DE gels of 7 days-old cultures of P. ostreatus (GIF 231 kb)

High resolution image (TIFF 848 kb)

Fig. S2

Raw images of 2-DE gels of 12 days-old cultures of P. ostreatus (GIF 260 kb)

High resolution image (TIFF 912 kb)

Fig. S3

Computerized image of protein maps of 7 days-old P. ostreatus cultures (“Master gel”). Statistically significant proteins are marked in red. (GIF 145 kb)

High resolution image (TIFF 94 kb)

Fig. S4

Computerized image of protein maps of 12 days-old P. ostreatus cultures (“Master gel”). Statistically significant proteins are marked in red. (GIF 147 kb)

High resolution image (TIFF 134 kb)

Tab. S1

Summary matching report for 2-DE gel comparisons. (PDF 195 kb)

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Petráčková, D., Halada, P., Bezoušková, S. et al. A two-dimensional protein map of Pleurotus ostreatus microsomes-proteome dynamics. Folia Microbiol 61, 63–71 (2016). https://doi.org/10.1007/s12223-015-0410-2

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  • DOI: https://doi.org/10.1007/s12223-015-0410-2

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