Kinetic Parameters and Cytotoxic Activity of Recombinant Methionine γ-Lyase from Clostridium tetani, Clostridium sporogenes, Porphyromonas gingivalis and Citrobacter freundii

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  • 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
  • Affiliations:
    1. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences
    2. Institute of Gene Biology, Russian Academy of Sciences
    3. Blokhin Cancer Research Center, Russian Academy of Medical Sciences
    4. Gamaleya Research Institute of Epidemiology and Microbiology,
  • Issue: Vol 5, No 3 (2013)
  • Pages: 92-98
  • Section: Research Articles
  • URL: http://actanaturae.ru/2075-8251/article/view/10590
  • DOI: https://doi.org/10.32607/20758251-2013-5-3-92-98
  • 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.


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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Copyright (c) 2013 Morozova E.A., Kulikova V.V., Yashin D.V., Anufrieva N.V., Anisimova N.Y., Revtovich S.V., Kotlov M.I., Belyi Y.F., Pokrovsky V.S., Demidkina T.V.

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