Kinetic Parameters and Cytotoxic Activity of Recombinant Methionine γ-Lyase from Clostridium tetani, Clostridium sporogenes, Porphyromonas gingivalis and Citrobacter freundii
- Authors: Morozova E.A.1, Kulikova V.V.1, Yashin D.V.2, Anufrieva N.V.1, Anisimova N.Y.3, Revtovich S.V.1, Kotlov M.I.4, Belyi Y.F.4, Pokrovsky V.S.3, Demidkina T.V.1
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Affiliations:
- Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
- Institute of Gene Biology, Russian Academy of Sciences
- Blokhin Cancer Research Center, Russian Academy of Medical Sciences
- Gamaleya Research Institute of Epidemiology and Microbiology,
- Issue: Vol 5, No 3 (2013)
- Pages: 92-98
- Section: Research Articles
- Submitted: 17.01.2020
- Published: 15.09.2013
- URL: https://actanaturae.ru/2075-8251/article/view/10590
- DOI: https://doi.org/10.32607/20758251-2013-5-3-92-98
- ID: 10590
Cite item
Abstract
The steady-state kinetic parameters of pyridoxal 5’-phosphate-dependent recombinant methionine γ-lyase from three pathogenic bacteria, Clostridium tetani, Clostridium sporogenes, and Porphyromonas gingivalis, were determined in β- and γ-elimination reactions. The enzyme from C. sporogenes is characterized by the highest catalytic efficiency in the γ-elimination reaction of L-methionine. It was demonstrated that the enzyme from these three sources exists as a tetramer. The N-terminal poly-histidine fragment of three recombinant enzymes influences their catalytic activity and facilitates the aggregation of monomers to yield dimeric forms under denaturing conditions. The cytotoxicity of methionine γ-lyase from C. sporogenes and C. tetani in comparison with Citrobacter freundii was evaluated using K562, PC-3, LnCap, MCF7, SKOV-3, and L5178y tumor cell lines. K562 (IC50=0.4-1.3 U/ml), PC-3 (IC50=0.1-0.4 U/ml), and MCF7 (IC50=0.04-3.2 U/ml) turned out to be the most sensitive cell lines.
About the authors
E. A. Morozova
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
Email: vadimpokrovsky@yandex.ru
Russian Federation
V. V. Kulikova
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
Email: vadimpokrovsky@yandex.ru
Russian Federation
D. V. Yashin
Institute of Gene Biology, Russian Academy of Sciences
Email: vadimpokrovsky@yandex.ru
Russian Federation
N. V. Anufrieva
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
Email: vadimpokrovsky@yandex.ru
Russian Federation
N. Y. Anisimova
Blokhin Cancer Research Center, Russian Academy of Medical Sciences
Email: vadimpokrovsky@yandex.ru
Russian Federation
S. V. Revtovich
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
Email: vadimpokrovsky@yandex.ru
Russian Federation
M. I. Kotlov
Gamaleya Research Institute of Epidemiology and Microbiology,
Email: vadimpokrovsky@yandex.ru
Russian Federation
Y. F. Belyi
Gamaleya Research Institute of Epidemiology and Microbiology,
Email: vadimpokrovsky@yandex.ru
Russian Federation
V. S. Pokrovsky
Blokhin Cancer Research Center, Russian Academy of Medical Sciences
Author for correspondence.
Email: vadimpokrovsky@yandex.ru
Russian Federation
T. V. Demidkina
Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
Email: tvd@eimb.ru
Russian Federation
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