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Physicochemical Characteristics of a Variant of Chaperon GroEL Apical Domain Designed to Enhance the Expression and Stability of Target Proteins

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Abstract

This work describes the properties of a new protein, a modification of GroEL apical domain designed to be a leader in fusion systems. This polypeptide leader demonstrates a high level of expression in a bacterial system; it is soluble and retains its solubility during standard biochemical manipulations. The secondary structure of the protein and its thermostability, as well as the protein solubility, were studied in a wide temperature range. To simplify the subsequent purification of the target protein, the possibility of its chemical cleavage from the fused protein by methionine residues with cyanogen bromide is provided.

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Funding

This work was carried out with the financial support of the Ministry of Science and Higher Education of the Russian Federation, project no. RFMEFI57517X0151.

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Authors and Affiliations

  1. People’s Friendship University of Russia, 117198, Moscow, Russia

    M. S. Yurkova, V. A. Zenin & A. N. Fedorov

  2. Fundamentals of Biotechnology Federal Research Center, Russian Academy of Sciences, 119071, Moscow, Russia

    K. A. Kurov, O. I. Savvin, M. S. Yurkova, V. A. Zenin & A. N. Fedorov

  3. Institute of Protein Research, Russian Academy of Sciences, 142290, Pushchino, Moscow oblast, Russia

    G. S. Nagibina & B. S. Melnik

Authors
  1. K. A. Kurov
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  2. O. I. Savvin
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  3. M. S. Yurkova
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  4. V. A. Zenin
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  5. G. S. Nagibina
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  6. B. S. Melnik
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  7. A. N. Fedorov
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Corresponding author

Correspondence to A. N. Fedorov.

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The authors declare that they have no conflicts of interest.

This article does not contain any studies involving animals or human participants performed by any of the authors.

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Translated by I. Gordon

Abbreviations: AEBSF, 4-(2-aminoethyl)benzenesulfonyl fluoride; IPTG, isopropyl-β-D-thiogalactopyranoside; LB medium, lysogenic broth medium; PAGE, polyacrylamide gel electrophoresis; PBS, phosphate-buffered saline.

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Kurov, K.A., Savvin, O.I., Yurkova, M.S. et al. Physicochemical Characteristics of a Variant of Chaperon GroEL Apical Domain Designed to Enhance the Expression and Stability of Target Proteins. Appl Biochem Microbiol 55, 765–770 (2019). https://doi.org/10.1134/S0003683819080088

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  • DOI: https://doi.org/10.1134/S0003683819080088

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