Biochemical Studies of Mitochondrial Malate: Quinone Oxidoreductase from Toxoplasma gondii.
Biochemical Studies of Mitochondrial Malate: Quinone Oxidoreductase from Toxoplasma gondii.
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DOI：
10.3390/ijms22157830
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发表时间：
2021-07-22
影响因子：
5.6
通讯作者：
Inaoka DK
中科院分区：
文献类型：
--
作者：
Acharjee R;Talaam KK;Hartuti ED;Matsuo Y;Sakura T;Gloria BM;Hidano S;Kido Y;Mori M;Shiomi K;Sekijima M;Nozaki T;Umeda K;Nishikawa Y;Hamano S;Kita K;Inaoka DK
弓形虫是一种引起弓形虫病的原生动物寄生虫，感染了全球近三分之一的人口。缺乏有效的药物和疫苗以及抗药性寄生虫的出现突出了开发新药的必要性。线粒体电子传递链（ETC）是T.刚地在顶复门寄生虫中，苹果酸：醌氧化还原酶（MQO）是属于ETC的单聚膜蛋白，也是三羧酸循环的关键成员，最近有人认为它在富马酸循环中发挥作用，而富马酸循环是胞质嘌呤所需的。补救途径。于T.弓形虫，一种推定的MQO（TgMQO）在速殖子和缓殖子阶段表达，并且被认为是潜在的药物靶标，因为其直向同源物在哺乳动物宿主中不保守。作为评估TgMQO作为候选药物靶点的第一步，在这项研究中，我们开发了一种新的TgMQO在FN102（DE3）TAO中的表达系统，该菌株缺乏呼吸道细胞色素并依赖于替代氧化酶。该系统允许，第一次，表达和纯化的线粒体MQO家族酶，这是用于稳态动力学和底物特异性分析。已知的唯一MQO抑制剂阿魏烯醇也抑制TgMQO，IC50为0.822 μM，与恶性疟原虫MQO相比，表现出不同的抑制动力学。此外，我们的分析表明，存在的第三个结合位点的阿魏烯醇，这是不同的泛醌和苹果酸盐的网站。
Toxoplasma gondii is a protozoan parasite that causes toxoplasmosis and infects almost one-third of the global human population. A lack of effective drugs and vaccines and the emergence of drug resistant parasites highlight the need for the development of new drugs. The mitochondrial electron transport chain (ETC) is an essential pathway for energy metabolism and the survival of T. gondii. In apicomplexan parasites, malate:quinone oxidoreductase (MQO) is a monotopic membrane protein belonging to the ETC and a key member of the tricarboxylic acid cycle, and has recently been suggested to play a role in the fumarate cycle, which is required for the cytosolic purine salvage pathway. In T. gondii, a putative MQO (TgMQO) is expressed in tachyzoite and bradyzoite stages and is considered to be a potential drug target since its orthologue is not conserved in mammalian hosts. As a first step towards the evaluation of TgMQO as a drug target candidate, in this study, we developed a new expression system for TgMQO in FN102(DE3)TAO, a strain deficient in respiratory cytochromes and dependent on an alternative oxidase. This system allowed, for the first time, the expression and purification of a mitochondrial MQO family enzyme, which was used for steady-state kinetics and substrate specificity analyses. Ferulenol, the only known MQO inhibitor, also inhibited TgMQO at IC50 of 0.822 μM, and displayed different inhibition kinetics compared to Plasmodium falciparum MQO. Furthermore, our analysis indicated the presence of a third binding site for ferulenol that is distinct from the ubiquinone and malate sites.
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弓形虫病发育中的药物：进步，挑战和当前状态。
DOI：
10.2147/dddt.s60973
发表时间：
2017
期刊：
Drug design, development and therapy
影响因子：
--
作者：
Alday PH;Doggett JS
通讯作者：
Doggett JS
影响因子：
4.3
作者：
Borisov, Vitaliy B.;Gennis, Robert B.;Hemp, James;Verkhovsky, Michael I.
通讯作者：
Verkhovsky, Michael I.
影响因子：
4.8
作者：
Bulusu, Vinay;Jayaraman, Vijay;Balaram, Hemalatha
通讯作者：
Balaram, Hemalatha
影响因子：
2.4
作者：
BUXTON, D;INNES, EA
通讯作者：
INNES, EA
影响因子：
--
作者：
Blume, Martin;Seeber, Frank
通讯作者：
Seeber, Frank