Suppression of telomerase activity in leukemic cells by mutant forms of Rhodospirillum rubrum L-asparaginase

Pokrovskaya, M. V.; Zhdanov, Dmitry; Eldarov, M. A.; Александрова, С. С.; Veselovsky, A. V.; Pokrovskiy, V. S.; Grishin, D. V.; Gladilina, Ju. A.; Sokolov, N. N.

doi:10.1134/s1990750817030088

Cited by 3 publications

(3 citation statements)

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“…Erwinia carotovora (Pectobacterium carotovorum) G281S Increased half-inactivation temperature; decreased catalytic activity for L-asparagine [76] Dickeya dadantii (Erwinia chrysanthemi) E63Q Decreased L-glutaminase activity [177] Dickeya dadantii (Erwinia chrysanthemi) D133V, D133L Higher thermal stability [73] Dickeya dadantii (Erwinia chrysanthemi) P285T Eight-fold reduced immunogenicity [99] Dickeya dadantii (Erwinia chrysanthemi) N41D, N281D Increased asparaginase activity and decreased L-glutaminase activity [81] Helicobacter pylori T16D Reduction of L-asparaginase and L-glutaminase activities [74] Helicobacter pylori Q63E Decreased glutaminase activity [74] Helicobacter pylori T95E Reduction of L-asparaginase and L-glutaminase activities [74] Helicobacter pylori M121C/T169M Undetectable L-glutaminase activity [133] Pyrococcus furiosus K274E Increased L-asparaginase activity, resistance to proteolytic digestion and no displayed glutaminase activity [80] Rhodospirillum rubrum D60K, F61L Improvement of kinetic parameters and enzyme stabilization [178] Rhodospirillum rubrum E149R, V150P, F151T Able to reduce the expression hTERT subunit of telomerase and suppress telomerase activity [24,25,163] Saccharomyces cerevisiae T64A, Y78A, T141A, K215A 99.9% loss of activity [179] Wolinella succinogenes V23Q, K24T Resistance to trypsin degradation, decreased glutaminase activity and reduced immunogenicity [170]…”

Section: Source Mutations Results Achievedmentioning

confidence: 99%

“…By site-directed mutagenesis, active and stable mutant forms of type I short-chain cytoplasmic L-ASNase from Rhodospirillum rubrum (RrA) were cloned and purified. The mutants containing the substitutions E149R, V150P, F151T, E149R and V150P were capable of reducing the expression of the hTERT (human telomerase reverse transcriptase) telomerase subunit, suppressing telomerase activity in cancer cell lines and affecting the growth of xenograft tumors [ 24 , 25 , 163 ]. This observation suggests that RrA targets tumor growth by a dual mechanism involving asparagine deprivation and telomerase inhibition.…”

Section: Alternative Approaches To the Development Of Antitumor L-asn...mentioning

confidence: 99%

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Molecular Analysis of L-Asparaginases for Clarification of the Mechanism of Action and Optimization of Pharmacological Functions

et al. 2022

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L-asparaginases (EC 3.5.1.1) are a family of enzymes that catalyze the hydrolysis of L-asparagine to L-aspartic acid and ammonia. These proteins with different biochemical, physicochemical and pharmacological properties are found in many organisms, including bacteria, fungi, algae, plants and mammals. To date, asparaginases from E. coli and Dickeya dadantii (formerly known as Erwinia chrysanthemi) are widely used in hematology for the treatment of lymphoblastic leukemias. However, their medical use is limited by side effects associated with the ability of these enzymes to hydrolyze L-glutamine, as well as the development of immune reactions. To solve these issues, gene-editing methods to introduce amino-acid substitutions of the enzyme are implemented. In this review, we focused on molecular analysis of the mechanism of enzyme action and to optimize the antitumor activity.

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Section: Source Mutations Results Achievedmentioning

confidence: 99%

Section: Alternative Approaches To the Development Of Antitumor L-asn...mentioning

confidence: 99%

Molecular Analysis of L-Asparaginases for Clarification of the Mechanism of Action and Optimization of Pharmacological Functions

et al. 2022

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“…Мутантная форма внутриклеточной L-аспарагиназы из Rhodospirillum rubrum RrA 17N, K149E проявляла цитотоксическую (антипролиферативную) активность in vitro на клетках Jurkat, HL-60, MOLT 4 и др. [12] и in vivo у мышей с лимфаденозом Фишера L5178y [13], а также подавляла на 80% экспрессию теломеразы in vitro [14]. Вариант RrА 17N, E149R, V150P с дополнительными заменами E149R и V150P еще сильнее снижал активность теломеразы до 15% от контроля в клетках Jurkat, HL-60, MOLT 4 и др.…”

Section: бактериальныеunclassified

Physical-Chemical Properties of L-asparaginase Mutants From Rhodospirillum Rubrum which Showed Antitelomerase Activity

Pokrovskaya

Александрова

Veselovsky

et al. 2019

BMCRM

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Rru_A3730 protein is a bacterial Rhodospirillum rubrum L-asparaginase (RrA), which is known by its anticancer activity. RrA variants with point amino acid substitutions in the region of 150 amino acids residues: RrA17N, K149E, RrAE149R, V150P, F151T, RrА17N, E149R, V150P, RrAE149R, V150P, showed antiproliferative properties, and also by their ability to suppress telomerase activity. This work is devoted to comparison of physical-chemical and catalytic properties of these mutant forms of RrA. It is shown that pH optimum is in the alkaline zone (8.5 – 9.3); L-glutaminase and D-asparaginase activity is respectively not more than 0.1% and 1.6% of L-asparaginase for all studied variants of RrA. The presence of the N17-terminal amino acid sequence MASMTGGQMGRGSSRQ of the capsid protein of bacteriophage T7 in the RrA structure leads to an increase in the thermal stability of mutant RrA analogues (from 50°C to 56°C) and their resistance to denaturation in the presence of 3 – 4 M urea. It is of Metal ions exhibit multidirectional effects on L-asparaginase activity of RrA. K+, Ca2+, Zn2+, Cs+, Co2+ in significantly affect the activity of L-asparaginase, while Mn2+, Cu2+, Fe3+ ions inhibit it. There was no correlation between antitelomerase (antiproliferative) activity and kinetic properties of mutant forms of L-asparaginase RrA.

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Improvement of Biocatalytic Properties and Cytotoxic Activity of L-Asparaginase from Rhodospirillum rubrum by Conjugation with Chitosan-Based Cationic Polyelectrolytes

et al. 2022

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L-asparaginases (L-ASNases, EC 3.5.1.1) are a family of enzymes that are widely used for the treatment of lymphoblastic leukemias. L-ASNase from Rhodospirillum rubrum (RrA) has a low molecular weight, low glutaminase activity, and low immunogenicity, making it a promising enzyme for antitumor drug development. In our work, the complex formation and covalent conjugation of the enzyme with synthetic or natural polycationic polymers was studied. Among non-covalent polyelectrolyte complexes (PEC), polyethyleneimine (PEI) yielded the highest effect on RrA, increasing its activity by 30%. The RrA-PEI complex had increased stability to trypsinolysis, with an inactivation constant decrease up to 10-fold compared to that of the native enzyme. The covalent conjugation of RrA with chitosan-PEI, chitosan-polyethylene glycol (chitosan-PEG), and chitosan-glycol resulted in an increase in the specific activity of L-asparagine (up to 30%). RrA-chitosan-PEG demonstrated dramatically (by 60%) increased cytotoxic activity for human chronic myeloma leukemia K562 cells in comparison to the native enzyme. The antiproliferative activity of RrA and its conjugates was significantly higher (up to 50%) than for that of the commercially available EcA at the same concentration. The results of this study demonstrated that RrA conjugates with polycations can become a promising strategy for antitumor drug development.

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Suppression of telomerase activity in leukemic cells by mutant forms of Rhodospirillum rubrum L-asparaginase

Cited by 3 publications

References 29 publications

Molecular Analysis of L-Asparaginases for Clarification of the Mechanism of Action and Optimization of Pharmacological Functions

Molecular Analysis of L-Asparaginases for Clarification of the Mechanism of Action and Optimization of Pharmacological Functions

Physical-Chemical Properties of L-asparaginase Mutants From Rhodospirillum Rubrum which Showed Antitelomerase Activity

Improvement of Biocatalytic Properties and Cytotoxic Activity of L-Asparaginase from Rhodospirillum rubrum by Conjugation with Chitosan-Based Cationic Polyelectrolytes

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