1. Institute of Biomedical Chemistry, Moscow, Russia 2. Research Center of Biotechnology RAS, Moscow, Russia 3. N.N. Blokhin Russian Cancer Research Center, Moscow, Russia
The active and stable mutant forms of short chain cytoplasmic L-asparaginase type I of Rhodospirillum rubrum (RrA): RrA+N17, D60K, F61L, RrA+N17, A64V, E67K, RrA+N17, E149R, V150P, RrAE149R, V150P and RrAE149R, V150P, F151T were obtained by the method of site-directed mutagenesis. It is established that variants RrA-N17, E149R, V150P, F151T and RrАE149R, V150P are capable to reduce an expression hTERT subunit of telomerase and, hence, activity of telomeres in Jurkat cells, but not in cellular lysates. During too time, L-asparaginasеs of Escherichia coli, Erwinia carotovora and Wolinella succinogenes, mutant forms RrА+N17, D60K, F61L and RrА+N17, A64V, E67K do not suppress of telomerase activity. The assumption of existence in structure RrA of areas (amino acids residues in the position 146-164, 1-17, 60-67) which are responsible for suppression of telomerase activity is made. The received results show that antineoplastic activity of some variants RrA is connected both with reduction of concentration of free L-asparagine, and with expression suppression of hTERT telomerase subunit, that opens new prospects for antineoplastic therapy.
Download PDF:
Keywords: L-asparaginase Rhodospirillum rubrum, antineoplastic effect of L-asparaginase, a site-directed mutagenesis, telomerase, hTERT subunit of telomerase
Citation:
Pokrovskaya M.V., Zhdanov D.D., Eldarov M.A., Aleksandrova S.S., Veselovskiy A.V., Pokrovskiy V.S., Grishin D.V., Gladilina Ju.A., Sokolov N.N. (2017) Suppression of telomerase activity leukemic cells by mutant forms of Rhodospirillum rubrum L-asparaginase. Biomeditsinskaya Khimiya, 63(1), 62-74.
Pokrovskaya M.V. et al. Suppression of telomerase activity leukemic cells by mutant forms of Rhodospirillum rubrum L-asparaginase // Biomeditsinskaya Khimiya. - 2017. - V. 63. -N 1. - P. 62-74.
Pokrovskaya M.V. et al., "Suppression of telomerase activity leukemic cells by mutant forms of Rhodospirillum rubrum L-asparaginase." Biomeditsinskaya Khimiya 63.1 (2017): 62-74.
Pokrovskaya, M. V., Zhdanov, D. D., Eldarov, M. A., Aleksandrova, S. S., Veselovskiy, A. V., Pokrovskiy, V. S., Grishin, D. V., Gladilina, Ju. A., Sokolov, N. N. (2017). Suppression of telomerase activity leukemic cells by mutant forms of Rhodospirillum rubrum L-asparaginase. Biomeditsinskaya Khimiya, 63(1), 62-74.
Horowitz B., Madras B.K., Meister A., Old L.J., Boyes E.A., Stockert E. (1968) Science, 160, 533-535. CrossRef Scholar google search
Ueno T., Ohtawa K., Mitsui K., Kodera Y., Hiroto M., Matsushima A., Inada Y., Nishimura H. (1997) Leukemia, 11, 1858-1861. CrossRef Scholar google search
Green M.R., Sambrook J., MacCallum P. (2012) Molecular cloning: a laboratory manual (4th Edn), Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press ISBN 978-1-936113-41-5. Scholar google search
Vinci S., Giannarini G., Selli C., Kuncova J., Villari D., Valent F., Orlando C. (2011) Urology Oncology, 29, 150-156. CrossRef Scholar google search
Branca M., Giorgi C., Ciotti M., Santini D., Di Bonito L., Costa S., Benedetto A., Bonifacio D, Di Bonito P., Paba P., Accardi L., Mariani L., Ruutu M., Syrjanen S., Favalli C., Syrjanen K. (2006) Diagnostic Cytopathology, 34, 739-748. CrossRef Scholar google search
