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

• Published in: Biomeditsinskaia khimiia Vol. 63; no. 1; p. 62
• Main Authors: 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
• Format: Journal Article
• Language: Russian
• Published: Russia (Federation) 01.01.2017
• Subjects: Amino Acid Sequence; Antineoplastic Agents - chemistry; Antineoplastic Agents - metabolism; Antineoplastic Agents - pharmacology; Asparaginase - chemistry; Asparaginase - genetics; Asparaginase - metabolism; Asparaginase - pharmacology; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacterial Proteins - pharmacology; Cell Line, Tumor; Cell Survival - drug effects; Escherichia coli - chemistry; Escherichia coli - enzymology; Escherichia coli - genetics; Gene Expression; HL-60 Cells; Humans; Jurkat Cells; Models, Molecular; Mutagenesis, Site-Directed; Pectobacterium carotovorum - chemistry; Pectobacterium carotovorum - enzymology; Pectobacterium carotovorum - genetics; Plasmids - chemistry; Plasmids - metabolism; Point Mutation; Protein Structure, Secondary; Rhodospirillum rubrum - chemistry; Rhodospirillum rubrum - enzymology; Rhodospirillum rubrum - genetics; Species Specificity; Structure-Activity Relationship; Telomerase - antagonists & inhibitors; Telomerase - genetics; Telomerase - metabolism; Telomere - chemistry; Telomere - drug effects; Wolinella - chemistry; Wolinella - enzymology; Wolinella - genetics; a site-directed mutagenesis; antineoplastic effect of L-asparaginase; hTERT subunit of telomerase; telomerase; L-asparaginase Rhodospirillum rubrum
• ISSN: 2310-6972
• DOI: 10.18097/PBMC20176301062

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-asparaginases 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.