Document Type : Original Article
Authors
1 Student Research Committee, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
2 Department of Biology, Faculty of Basic Sciences, University of Maragheh, Maragheh, Iran
3 Department of Medical Genetics and Pathology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
4 Department of Anatomical Sciences, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran
5 Department of Medical Laboratory Sciences, School of Paramedical Sciences, Ardabil University of Medical Sciences, Ardabil, Iran
Abstract
Background: The L-asparaginase enzyme is used as an anticancer agent in treating acute lymphoblastic leukemia (ALL). Moreover, it has widespread applications in medicine, food, and pharmaceutical industries.
Methods: The nucleotide and amino acid sequences of L-asparaginase derived from Bacillus subtilis BEST7613, Kibdelosporangium sp. MJ126-NF4, and Thermococcus kodakarensis KOD1 have been obtained from the GenBank and the NCBI databases. The EMBOSS Water pairwise sequence alignments were performed using ClustalW 1.83. Prediction of secondary and tertiary protein structures of the different L-asparaginase molecules studied was done using SWISS-MODEL software. In addition, the protein domains of L-asparaginase originating from the three mentioned bacteria were analyzed using PROSITE software. Theoretical isoelectric point (pI), molecular weight, and amino acid composition were predicted using the protein pI calculator (http://isoelectric.ovh.org/).
Results: Despite the structural differences in L-asparaginase enzymes in the three bacterial strains, there were no differences in their functional characteristics, including molecular weight, pI, and functional domain.
Conclusion: Analyzing structural differences and finding functional similarities can be useful in designing drugs with higher stability and biological half-life. Our analysis showed that proteins with different structures could have similar functional characteristics, which proves the codon usage hypothesis.
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Main Subjects
- Vardiman JW, Harris NL, Brunning RD. The World Health Organization (WHO) classification of the myeloid neoplasms. Blood. 2002;100(7):2292-302. doi: 10.1182/ blood-2002-04-1199.
- Hijiya N, van der Sluis IM. Asparaginase-associated toxicity in children with acute lymphoblastic leukemia. Leuk Lymphoma. 2016;57(4):748-57. doi: 10.3109/10428194.2015.1101098.
- Ammulu MA, Rosaiah G, Managmauri UK, Vemuri PK, Rao KR, Sudhakar P. L-asparaginase a biotherapeutic for acute lymphoblastic leukemia-a molecular perspective. Curr Trends Biotechnol Pharm. 2017;11(4):424-43. doi: 10.6084/ m9.figshare.13297637.
- Phan LM, Yeung SC, Lee MH. Cancer metabolic reprogramming: importance, main features, and potentials for precise targeted anti-cancer therapies. Cancer Biol Med. 2014;11(1):1-19. doi: 10.7497/j.issn.2095-3941.2014.01.001.
- Takahashi H, Inoue J, Sakaguchi K, Takagi M, Mizutani S, Inazawa J. Autophagy is required for cell survival under L-asparaginase-induced metabolic stress in acute lymphoblastic leukemia cells. Oncogene. 2017;36(30):4267- 76. doi: 10.1038/onc.2017.59.
- Kelo E, Noronkoski T, Mononen I. Depletion of L-asparagine supply and apoptosis of leukemia cells induced by human glycosylasparaginase. Leukemia. 2009;23(6):1167-71. doi: 10.1038/leu.2008.387.
- Vimal A, Kumar A. Biotechnological production and practical application of L-asparaginase enzyme. Biotechnol Genet Eng Rev. 2017;33(1):40-61. doi: 10.1080/02648725.2017.1357294.
- Alam S, Pranaw K, Tiwari R, Khare SK. Recent development in the uses of asparaginase as food enzyme. In: Parameswaran B, Varjani S, Raveendran S, eds. Green Bio-processes: Enzymes in Industrial Food Processing. Singapore: Springer; 2019. p. 55-81. doi: 10.1007/978-981-13-3263-0_5.
- Jia R, Wan X, Geng X, Xue D, Xie Z, Chen C. Microbial L-asparaginase for application in acrylamide mitigation from food: current research status and future perspectives. Microorganisms. 2021;9(8):1659. doi: 10.3390/ microorganisms9081659.
- Cachumba JJ, Antunes FA, Peres GF, Brumano LP, Santos JC, Da Silva SS. Current applications and different approaches for microbial L-asparaginase production. Braz J Microbiol. 2016;47 Suppl 1(Suppl 1):77-85. doi: 10.1016/j. bjm.2016.10.004.
- Krishnapura PR, Belur PD, Subramanya S. A critical review on properties and applications of microbial L-asparaginases. Crit Rev Microbiol. 2016;42(5):720-37. doi: 10.3109/1040841x.2015.1022505.
- Quax TE, Claassens NJ, Söll D, van der Oost J. Codon bias as a means to fine-tune gene expression. Mol Cell. 2015;59(2):149- 61. doi: 10.1016/j.molcel.2015.05.035.
- Kumar K, Kaur J, Walia S, Pathak T, Aggarwal D. L-asparaginase: an effective agent in the treatment of acute lymphoblastic leukemia. Leuk Lymphoma. 2014;55(2):256-62. doi: 10.3109/10428194.2013.803224.
- Gurung N, Ray S, Bose S, Rai V. A broader view: microbial enzymes and their relevance in industries, medicine, and beyond. Biomed Res Int. 2013;2013:329121. doi: 10.1155/2013/329121.
- van Vught R, Blenke EO, Mastrobattista E, Martin N, Roovers R, van Beijeren Bergen En Henegouwen P, et al. Incorporation of unnatural amino acids in nanobodies. In: Functionalized Nanobodies for Cancer Therapy. Utrecht University Repository; 2014. p. 97.
- Maggi M, Mittelman SD, Parmentier JH, Colombo G, Meli M, Whitmire JM, et al. A protease-resistant Escherichia coli asparaginase with outstanding stability and enhanced antileukaemic activity in vitro. Sci Rep. 2017;7(1):14479. doi: 10.1038/s41598-017-15075-4.
- Patel N, Krishnan S, Offman MN, Krol M, Moss CX, Leighton C, et al. A dyad of lymphoblastic lysosomal cysteine proteases degrades the antileukemic drug L-asparaginase. J Clin Invest. 2009;119(7):1964-73. doi: 10.1172/jci37977.
- Rutter DA, Wade HE. The influence of the iso-electric point of L-asparaginase upon its persistence in the blood. Br J Exp Pathol. 1971;52(6):610-4.
- Vrkljan M, Foster TM, Powers ME, Henkin J, Porter WR, Staack H, et al. Thermal stability of low molecular weight urokinase during heat treatment. II. Effect of polymeric additives. Pharm Res. 1994;11(7):1004-8. doi: 10.1023/a:1018935420680.
- Horovitz A, Matthews JM, Fersht AR. Alpha-helix stability in proteins. II. Factors that influence stability at an internal position. J Mol Biol. 1992;227(2):560-8. doi: 10.1016/0022- 2836(92)90907-2.
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