Construction and Characterization of a Recombinant Mutant Homolog of the CheW Protein from Thermotoga petrophila RKU-1
- Authors: Grishin D.V.1, Zhdanov D.D.1, Gladilina J.A.1, Pokrovsky V.S.1, Podobed O.V.1, Pokrovskaya M.V.1, Aleksandrova S.S.1, Milyushkina A.L.1, Vigovskiy M.A.1, Sokolov N.N.1
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Affiliations:
- Institute of Biomedical Chemistry
- Issue: Vol 12, No 2 (2018)
- Pages: 143-150
- Section: Article
- URL: https://journals.rcsi.science/1990-7508/article/view/198179
- DOI: https://doi.org/10.1134/S1990750818020051
- ID: 198179
Cite item
Abstract
In this work, a recombinant chemotaxis CheW protein from Thermotoga petrophila RKU-1 (TpeCheW) and its mutant homologue (TpeCheW-mut) have been obtained. Despite the low homology with the CheW protein of the Escherichia coli bacteria, these proteins do not cause metabolic overload and are well expressed by E. coli laboratory strains. A wide range of features and parameters important for isolation of the TpeCheW-mut protein, such as stability over a wide range of temperatures and pH values, high level of expression, solubility, and the possibility of using simple low-stage purification schemes, including heat pretreatment, has been recognized. Possible directions of using this protein in the practice of scientific and applied research have been formulated and justified.
About the authors
D. V. Grishin
Institute of Biomedical Chemistry
Author for correspondence.
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121
D. D. Zhdanov
Institute of Biomedical Chemistry
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121
Ju. A. Gladilina
Institute of Biomedical Chemistry
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121
V. S. Pokrovsky
Institute of Biomedical Chemistry
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121
O. V. Podobed
Institute of Biomedical Chemistry
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121
M. V. Pokrovskaya
Institute of Biomedical Chemistry
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121
S. S. Aleksandrova
Institute of Biomedical Chemistry
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121
A. L. Milyushkina
Institute of Biomedical Chemistry
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121
M. A. Vigovskiy
Institute of Biomedical Chemistry
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121
N. N. Sokolov
Institute of Biomedical Chemistry
Email: molbiol_ibm@inbox.ru
Russian Federation, ul. Pogodinskaya 10, Moscow, 119121
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