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

E. A. Morozova; Морозова E. A.; V. V. Kulikova; Куликова В. В.; D. V. Yashin; Яшин Д. В.; N. V. Anufrieva; Ануфриева Н. В.; N. Y. Anisimova; Анисимова Н. Ю.; S. V. Revtovich; Ревтович С. В.; M. I. Kotlov; Котлов M. И.; Y. F. Belyi; Белый Ю. Ф.; V. S. Pokrovsky; Покровский В. С.; T. V. Demidkina; Демидкина T. В.

doi:10.32607/20758251-2013-5-3-92-98

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
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
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

Full Text

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.

Keywords

kinetic parameters, methionine γ-lyase, pathogenic microorganisms, oligomeric structure, cytotoxicity

References

Tanaka H., Esaki N., Soda K. // EnzymeMicrob. Technol. 1985, V.7, P.530-537
El-Sayed A.S. // Appl. Microbiol. Biotechnol. 2010, V.86, P.445-467
Nakayama T., Esaki N., Lee W.-J., Tanaka I., Tanaka H., Soda K. // Agric. Biol. Chem. 1984, V.48, P.2367-2369
Kreis W., Hession C. // Cancer Research 1973, V.33, P.1862-1865
Yoshimura M., Nakano Y., Yamashita Y., Oho T., Saito T., Koga T. // Infection Immunity. 2000, V.68, P.6912-6916
Tokoro M., Asai T., Kobayashi S., Takeuchi T., Nozaki T. // J. Biol. Chem. 2003, V.278, P.42717-42727
Lockwood B., Coombs G. // Biochem. J. 1991, V.279, P.675-682
Cellarier E., Durando X., Vasson M.P., Farges M.C., Demiden A., Maurizis J.C., Madelmont J.C., Chollet P. // Cancer Treat Rev. 2003, V.29, P.488-489
Yoshioka T., Wada T., Uchida N., Maki H., Yoshida H., Ide N., Kasai H., Hojo K., Shono K., Maekawa R. // Cancer Research 1998, V.58, P.2583-2587
Miki K., Xu M., An Z., Wang X., Yang M., Al-Refaie W., Sun X., Baranov E., Tan Y., Chishima T. // Cancer Gene Ther. 2000, V.7, P.332-338
Miki K., Al-Refaie W., Xu M., Jiang P., TanY. I.O., Bouvet M., Zhao M., Gupta A., Chishima T., Shimada H. // Cancer Research 2000, V.60, P.2696-2702
Tan Y., Xu M., Hoffman R.M. // Anticancer Res. 2010, V.30, P.1041-1046
El-Sayed A.S., Shouman S.A., Nassrat H.M. // Enzyme Microb. Technol. 2012, V.51, P.200-210
Revtovich S.V., Morozova Е.А., Anufrieva N.V., Kotlov M.I., Belyi Y.F., Demidkina T.V. // Dokl. Biochem. Biophys. 2012, V.445, P.187-193
Studier F.W. // ProteinExpr. Purif. 2005, V.41, P.207-234
Manukhov I.V., Mamaeva D.V., Morozova E.A., Rastorguev S.M., Faleev N.G., Demidkina T.V., Zavilgelsky G.B. // Biochemistry (Mosc). 2006, V.71, P.454-463
Morozova E.A., Bazhulina N.P., Anufrieva N.V., Mamaeva D.V., Tkachev Y.V., Streltsov S.A., Timofeev V.P., Faleev N.G., Demidkina T.V. // Biochemistry (Mosc). 201, V.75, P.1435-1445
Laemmli U.K. // Nature 1970, V.227, P.680-685
Meyer T.S., Lamberts B.L. // Biochim. Biophys. Acta . 1965, V.107, P.144-145
Towbin H., Staehelin T., Gordon J. // Proc.Natl.Acad.Sci. USA. 1979, V.76, P.4350-4354
Mossman T. // J. Immunol. Methods. 1983, V.65, P.55-63
Xu W., Zhang X., Qian H., Zhu W., Sun X., Hu J., Zhou H., Chen Y. // Exp.Biol.Med. 2004, V.229, P.623-631
Tan Y., Xu M., Hoffman R.M. // Anticancer Res. 2010, V.30, P.141-1046
Kudou D., Misaki S., Yamashita M., Tamura T., Esaki N., Inagaki K. // Biosci. Biotechnol. Biochem. 2008, V.72, P.1722-1730
Nakayama T., Esaki N., Sugie K., Berezov T.T., Tanaka H., Soda K. // Anal. Biochem. 1884, V.138, P.421-424
Mamaeva D.V., Morozova E.A., Nikulin A.D., Revtovich S.V., Nikonov S.V., Garber M.B., Demidkina T.V. // Acta Cryst. 2005, V.61, P.546-549
Chant A., Kraemer-Pecore C.M., Watkin R., Kneale G.G. // Protein Expr. Purif. 2005, V.39, P.152-159
Wu J., Filutowicz M. // Acta Biochem. Pol. 1999, V.46, P.591-599
Esteban-Torres M., Alvarez Y., Acebrón I., de las Rivas B., Muñoz R., Kohring G.W., Roa A.M., Sobrino M., Mancheño J.M. // FEBS Lett. 2012, V.586, P.3127-3133
Pokrovskaya M.V., Aleksandrova S.S., Pokrovsky V.S., Omeljanjuk N.M., Borisova A.A., Anisimova N.Yu., Sokolov N.N. // Protein Expr. Purif. 2012, V.82, P.150-154
Sokolov N.N., Zanin V.A., Aleksandrova S.S. // Aspects of medicinal chemistry // Vopr. Med. Khim. 2000, V.46, P.531-548
Krasotkina Y.V., Gladilina Y.A., Borisova А.А., Gervaziev Y.V., Abakumova О.Y., Zanin V.А., Sokolov N.N. // Aspects of biological, medical and pharmaceutical chemistry // Vopr. Biol. Med. Pharm. Chem. 2008, №3, P.18-21

Supplementary files

Supplementary Files

Action

1. JATS XML

Download

Username
Password
Remember me

Forgot password?	Register

Username
Password
Remember me

Forgot password?	Register

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

Full Text

Abstract

Keywords

About the authors

E. A. Morozova

V. V. Kulikova

D. V. Yashin

N. V. Anufrieva

N. Y. Anisimova

S. V. Revtovich

M. I. Kotlov

Y. F. Belyi

V. S. Pokrovsky

T. V. Demidkina

References

Supplementary files

This website uses cookies