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

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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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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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