Kinetic Study of Plasmodium falciparum Orotate Phosphoribosyl Transferase in the Presence of Activator and Inhibitor
การศึกษาจลนศาสตร์ของเอนไซม์ออโรเทต ฟอสฟอไรโบซิลทรานเฟอเรสของเชื้อพลาสโมเดียม ฟัลซิพารัมในสภาวะที่มีตัวกระตุ้นและตัวยับยั้ง
Keywords:
Plasmodium falciparum , orotate phosphoribosyl transferase , activation, inhibition, metal ionAbstract
Objective: To find the chemicals which have the stimulating or inhibitory properties on orotate phosphoribosyl transferase enzymes (OPRT) of Plasmodium falciparum. Method: construction of PfOPRT gene was ligated in pQE 30Xa plasmid and transfer into E. coli M15 and prove the construct recombinant plasmid. After it was proved to be the desired gene, the enzyme was expressed and purified. The performance was tested and metal ions were tested for activation and Orotidine 5'-Monophosphate (OMP) reaction products for inhibition. Result: The effective cofactor of OPRT is Mg2 + with Vmax = 64.25 µmol/min and Km = 58.82 µM and the enzyme activity without a cofactor is equal to zero. The cofactor was converted to Zn2 +, the OPRT activity was reduced was show Vmax = 9.35 µmol/min and Km= 136.24 µM. A reverse inhibition test was performed using the product obtained from the reaction OMP, resulting in a competitive inhibition of OPRT.
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References
World malaria report 2022. Geneva: World Health Organization. 2022.
Snow RW, Guerra CA, Noor AM, Myint HY, Hay SI. The global distribution of clinical episodes of Plasmodium falciparum malaria. Nature 2005;434(7030):214-7.
Dondorp AM, Nosten F, Yi P, Das D, Phyo AP, Tarning J, et al. Artemisinin resistance in Plasmodium falciparum malaria. The New England journal of medicine 2009;361(5):455-67.
Ridley RG. Medical need, scientific opportunity and the drive for antimalarial drugs. Nature 2002;415(6872):686-93.
White NJ. Antimalarial drug resistance. The Journal of clinical investigation 2004;113(8):1084-92.
Schwartz L, Brown GV, Genton B, Moorthy VS. A review of malaria vaccine clinical projects based on the WHO rainbow table. Malaria journal 2012;11:11.
Gardner MJ, Hall N, Fung E, White O, Berriman M, Hyman RW, et al. Genome sequence of the human malaria parasite Plasmodium falciparum. Nature 2002;419(6906):498-511.
Hay SI, Okiro EA, Gething PW, Patil AP, Tatem AJ, Guerra CA, et al. Estimating the global clinical burden of Plasmodium falciparum malaria in 2007. PLoS medicine 2010;7(6):e1000290.
Burton J. Bogitsh, & CEC, Oeltmann TN. Human Parasitology. New York: Elsevier; 2005.
Bray RS, Garnham PC. The life-cycle of primate malaria parasites. British medical bulletin 1982;38(2):117-22.
Markell E, Voge M. Medical parasitology. 4 ed. Philadelphia: WB Saunders; 1976.
Frugier M, Bour T, Ayach M, Santos MA, Rudinger-Thirion J, Théobald-Dietrich A, et al. Low Complexity Regions behave as tRNA sponges to help co-translational folding of plasmodial proteins. FEBS letters 2010;584(2):448-54.
Gero AM, O'Sullivan WJ. Purines and pyrimidines in malarial parasites. Blood cells 1990;16(2-3):467-84.
Krungkrai J, Cerami A, Henderson GB. Pyrimidine biosynthesis in parasitic protozoa: purification of a monofunctional dihydroorotase from Plasmodium berghei and Crithidia fasciculata. Biochemistry 1990;29(26):6270-5.
Reyes P, Rathod PK, Sanchez DJ, Mrema JE, Rieckmann KH, Heidrich HG. Enzymes of purine and pyrimidine metabolism from the human malaria parasite, Plasmodium falciparum. Molecular and biochemical parasitology 1982;5(5):275-90.
Huang M, Graves LM. De novo synthesis of pyrimidine nucleotides; emergin emerging interfaces with signal transduction pathways. Cellular and molecular life sciences: CMLS 2003;60(2):321-36.
Krungkrai SR, Prapunwattana P, Horii T, Krungkrai J. Orotate phosphoribosyl transferase and orotidine 5'-monophosphate decarboxylase exist as multienzyme complex in human malaria parasite Plasmodium falciparum Biochemical and biophysical research communications 2004;318(4):1012-8.
Krungkrai SR, Aoki S, Palacpac NM, Sato D, Mitamura T, Krungkrai J, et al. Human malaria parasite orotate phosphoribosyltransferase: functional expression, characterization of kinetic reaction mechanism and inhibition profile. Molecular and biochemical parasitology 2004;134(2):245-55.
Krungkrai SR, DelFraino BJ, Smiley JA, Prapunwattana P, Mitamura T, Horii T, et al. A novel enzyme complex of orotate phosphoribosyltransferase and orotidine 5'-monophosphate decarboxylase in human malaria parasite Plasmodium falciparum: physical association, kinetics, and inhibition characterization. Biochemistry 2005;44(5):1643-52.
Kanchanaphum P, Krungkrai J. Kinetic benefits and thermal stability of orotate phosphoribosyl transferase and orotidine 5'-monophosphate decarboxylase enzyme complex in human malaria parasite Plasmodium falciparum. Biochem Biophys Res Commun 2009;390:337-41.
Paojinda P, Imprasittichai W, Krungkrai SR, Palacpac NMQ, Horii T, Krungkrai J. Bifunctional activity of fused Plasmodium falciparum orotate phosphoribosyl transferase and orotidine 5′-monophosphate decarboxylase. Parasitol Int 2018;67(1):79-84.
Imprasittichai W, Krungkrai SR, Krungkrai J, Paojinda P. Expression and enzyme kinetic of fused Plasmodium falciparum orotate phosphoribosyl transferase and orotidine 5′-monophosphate decarboxylase in different Escherichia coli. Open Biochem. J 2023;17:1-8.
Imprasittichai W, Roytrakul S, Krungkrai SR, Krungkrai J. A unique insertion of low complexity amino acid sequence underlines protein-protein interaction in human malaria parasite orotate phosphoribosyltransferase and orotidine 5'-monophosphate decarboxylase. Asian Pac J Trop Med 2014;7:184-92.
Krungkrai J, Prapunwatana P, Wichitkul C, Reungprapavut S, Krungkrai SR, Horii T. Molecular biology and biochemistry of malarial parasite pyrimidine biosynthetic pathway. The Southeast Asian journal of tropical medicine and public health 2003;34:32-43.
Bhatia MB, Grubmeyer C. The Role of Divalent Magnesium in Activating the Reaction Catalyzed by Orotate Phosphoribosyltransferase. Archives of Biochemistry and Biophysics 1993;303(2):321–5.
Victor J, Leo-Mensah A, Sloan DL. Divalent metal ion activation of the yeast orotate phosphoribosyltransferase catalyzed reaction. Biochemistry 1979;18(16):3597-604.
Gonzalez-Segura L, Witte JF, McClard RW, Hurley TD. Ternary Complex Formation and Induced Asymmetry in Orotate Phosphoribosyltransferase. Biochemistry 2007;46(49):14075-86.
Donini S, Ferraris DM, Miggiano R, Massarotti A, Rizzi M. Structural investigations on orotate phosphoribosyltransferase from Mycobacterium tuberculosis, a key enzyme of the de novo pyrimidine biosynthesis. Scientific reports 2017;7(1):1-11.
Rathod PK, Leffers NP, Young RD. Molecular targets of 5-fluoroorotate in the human malaria parasite, Plasmodium falciparum. Antimicrobial agents and chemotherapy 1992;36(4):704-11.
Scott HV, Gero AM, O'Sullivan WJ. In vitro inhibition of Plasmodium falciparum by pyrazofurin, an inhibitor of pyrimidine biosynthesis de novo. Molecular and biochemical parasitology 1986;18(1):3-15.
Guerin PJ, Olliaro P, Nosten F, Druilhe P, Laxminarayan R, Binka F, et al. Malaria: current status of control, diagnosis, treatment, and a proposed agenda for research and development. The Lancet Infectious diseases 2002;2(9):564-73.
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