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GroEL Mediates Folding of Bacillus anthracis Serine/Threonine Protein Kinase, PrkC

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Abstract

Bacillus anthracis causes anthrax in human and animals. Both, signaling system such as two component system and endogenous chaperone system such as GroEL–GroES help bacteria to cope with the environmental challenges. Such molecular chaperones are the stress induced proteins that help bacteria to override unfavorable conditions by their moonlighting functions. Previous reports showed that PrkC and PrpC, the Ser/Thr kinase–phosphatase pair in B. anthracis, control phosphorylation of GroEL and regulate biofilm formation. In this study, we show that GroEL is involved in the folding of PrkC to active form. The proteins (GroEL, PrkC and PrpC) were expressed and purified by affinity chromatography. Purified GroEL was used for refolding of denatured PrkC and PrpC and observed that GroEL refolds PrkC but not PrpC as measured by their enzymatic activity. We also observed that purification of GroEL with six histidine tag using Cobalt-Agarose resin yielded superior quality GroEL protein with negligible contamination of non-specific proteins. Thus, cobalt resin can be a better choice for purification of many histidine tagged proteins, where Ni-NTA does not work very well.

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Acknowledgements

We thank Dr. Gunjan Arora for his valuable suggestions.

Funding

CSIR senior research fellowship to R.V. is acknowledged.

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Authors and Affiliations

  1. Delhi Technological University, New Delhi, Delhi, 110042, India

    Richa Virmani & Yasha Hasija

  2. Department of Zoology, University of Delhi, New Delhi, Delhi, 110007, India

    Richa Virmani & Yogendra Singh

Authors
  1. Richa Virmani
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  2. Yogendra Singh
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  3. Yasha Hasija
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Contributions

RV conceived and designed the experiments. YH and YS contributed materials, reagents and analysis tools. All authors have read and approved the manuscript.

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Correspondence to Yogendra Singh or Yasha Hasija.

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The authors declare that they have no competing interests.

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Virmani, R., Singh, Y. & Hasija, Y. GroEL Mediates Folding of Bacillus anthracis Serine/Threonine Protein Kinase, PrkC. Indian J Microbiol 58, 520–524 (2018). https://doi.org/10.1007/s12088-018-0744-y

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