Characterization of Toxoplasma gondii glyoxalase 1 and evaluation of inhibitory effects of curcumin on the enzyme and parasite cultures.
Characterization of Toxoplasma gondii glyoxalase 1 and evaluation of inhibitory effects of curcumin on the enzyme and parasite cultures.
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DOI：
10.1186/s13071-015-1268-5
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发表时间：
2015-12-23
影响因子：
3.2
通讯作者：
Xuan X
中科院分区：
文献类型：
--
作者：
Goo YK;Yamagishi J;Ueno A;Terkawi MA;Aboge GO;Kwak D;Hong Y;Chung DI;Igarashi M;Nishikawa Y;Xuan X
谷胱甘肽酶途径包括两种酶，谷胱甘肽酶1和2（Glo 1和Glo 2），是一种普遍存在的细胞系统，负责清除糖酵解过程中产生的细胞毒性甲基乙二醛。弓形虫（Toxoplasma gondii，T. gondii）速殖子，由于增加的糖酵解通量而产生甲基乙二醛。Glo 1抑制剂如姜黄素可被认为是抗原生动物、抗炎和抗癌治疗的候选药物。霸王克隆了弓形虫Glo 1基因（TgGlo 1），并制备了重组蛋白。通过向反应混合物中加入甲基乙二醛和谷胱甘肽来评价TgGlo 1和五种突变体的酶动力学。最后，利用体外培养的T.刚地成功构建了具有活性的重组TgGlo 1，并通过点突变验证了TgGlo 1的活性位点（E166和E251）。姜黄素在试验剂量下抑制重组TgGlo 1的酶活性以及体外培养的T.刚地Ki和IC_（50）分别为12.9 ± 0.5 μM和38.3 ± 0.9 μM。研究了姜黄素对TgGlo 1酶活性和寄生性T.弓形虫可用于开发治疗弓形虫病的有效药物。然而，考虑到已知姜黄素在微摩尔范围内对其他分子具有多种作用的事实，进一步阐明姜黄素对T.需要弓形虫。本文的在线版本（doi：10.1186/s13071-015-1268-5）包含补充材料，可供授权用户使用。
The glyoxalase pathway, which includes two enzymes, glyoxalase 1 and 2 (Glo1 and Glo2), is a ubiquitous cellular system responsible for the removal of cytotoxic methylglyoxal produced during glycolysis. Protozoan parasites, including Toxoplasma gondii (T. gondii) tachyzoites, produce methylglyoxal because of increased glycolytic fluxes. A Glo1 inhibitor such as curcumin could be considered a drug candidate for anti-protozoan, anti-inflammatory, and anti-cancer therapy. The T. gondii Glo1 gene (TgGlo1) was cloned and the recombinant protein was produced. Enzyme kinetics of TgGlo1 and five mutants were evaluated by adding methylglyoxal and glutathione to a reaction mixture. Finally, the inhibitory effects of various concentrations of curcumin on recombinant TgGlo1 were evaluated using in vitro cultures of T. gondii. Active recombinant TgGlo1 was successfully produced and the active sites (E166 and E251) of TgGlo1 were verified by point mutagenesis. Curcumin at the tested doses inhibited the enzymatic activity of recombinant TgGlo1 as well as the parasitic propagation of in vitro-cultured T. gondii. The Ki and IC50 were 12.9 ± 0.5 μM and 38.3 ± 0.9 μM, respectively. The inhibitory effect of curcumin on the enzymatic activity of TgGlo1 and parasitic propagation of T. gondii could be explored in the potential development of a potent drug for the treatment of toxoplasmosis. However, considering the fact that curcumin is known to have many effects on other molecules in the micromolar range, further elucidation of curcumin’s direct inhibition of the glyoxalase system of T. gondii will be needed. The online version of this article (doi:10.1186/s13071-015-1268-5) contains supplementary material, which is available to authorized users.