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Intracellular Proteomic Analysis of Streptomyces sp. MC1 When Exposed to Cr(VI) by Gel-Based and Gel-Free Methods

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Abstract

The actinobacterium Streptomyces sp. MC1 has previously shown the capacity to resist and remove Cr(VI) from liquid culture media. The aim of this work is to analyze the differential expression pattern of intracellular proteins when Streptomyces sp. MC1 is exposed to Cr(VI) in order to explain the molecular mechanisms of resistance that this microorganism possesses. For this purpose, 2D-PAGE and shotgun proteomic analyses (2D-nanoUPLC-ESI-MS/MS) were applied. The presence of Cr(VI) induced the expression of proteins involved in molecular biosynthesis and energy generation, chaperones with a key role in the repair of misfolded proteins and stress response, transcription proteins, proteins of importance in the DNA supercoiling, repair and replication, and dehydrogenases involved in oxidation–reduction processes. These dehydrogenases can be associated with the reduction of Cr(VI) to Cr(III). The results of this study show that proteins from the groups mentioned before are important to face the stress caused by the Cr(VI) presence and help the microorganism to counteract the toxicity of the metal. The use of two proteomic approaches resulted in a larger number of peptides identified, which is also transduced in a significant number of protein ID. This decreased the potential complexity of the sample because of the protein dynamic range, as well as increased the recovery of peptides from the gel after digestion.

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Acknowledgements

The authors thank the financial assistance of the National Agency for Scientific and Technological Promotion, Argentina (PICT 2013 No. 3170 to Dr. Villegas). The authors would also like to thank the English Scientific Writing Advice Group (GAECI) of the National University of San Luis for the revision of this article, and Mr. Doug Jennewein from USD-IT Research Computing for his help in the database installation and servers operation. Bonilla JO thanks CONICET for the awarded doctoral fellowship.

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Authors and Affiliations

  1. Instituto de Química San Luis (INQUISAL), CONICET, Chacabuco 917, 5700, San Luis, Argentina

    José O. Bonilla & Liliana B. Villegas

  2. Facultad de Química, Bioquímica y Farmacia, Universidad Nacional de San Luis, Chacabuco 917, 5700, San Luis, Argentina

    José O. Bonilla & Liliana B. Villegas

  3. Division of Basic Biomedical Sciences Sanford School of Medicine, University of South Dakota, Vermillion, SD, USA

    Eduardo A. Callegari

  4. Planta Piloto de Procesos Industriales Microbiológicos (PROIMI), CONICET, Av. Belgrano y Pje. Caseros, 4000, Tucumán, Argentina

    María C. Estevéz

  5. Facultad de Bioquímica, Química y Farmacia, Universidad Nacional de Tucumán, Ayacucho 471, 4000, Tucumán, Argentina

    María C. Estevéz

Authors
  1. José O. Bonilla
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  2. Eduardo A. Callegari
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  3. María C. Estevéz
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  4. Liliana B. Villegas
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Contributions

All authors contributed to the study conception and design of the work. Material preparation was performed by JB; electrophoresis assays were performed by JB, CE and LV; proteomic analyses were carried out by EC. Data collection and analysis were performed by JB, EC, and LV. The first draft of the manuscript was written by José Bonilla and the other authors read and modified the manuscript. All authors approved the final manuscript.

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Correspondence to Liliana B. Villegas.

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Bonilla, J.O., Callegari, E.A., Estevéz, M.C. et al. Intracellular Proteomic Analysis of Streptomyces sp. MC1 When Exposed to Cr(VI) by Gel-Based and Gel-Free Methods. Curr Microbiol 77, 62–70 (2020). https://doi.org/10.1007/s00284-019-01790-w

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