当前位置：
X-MOL 学术
›
ACS Infect. Dis.
›
论文详情
Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
FRET Flow Cytometry-Based High Throughput Screening Assay To Identify Disrupters of Glucose Levels in Trypanosoma brucei
ACS Infectious Diseases ( IF 4.0 ) Pub Date : 2018-05-09 00:00:00 , DOI: 10.1021/acsinfecdis.8b00058 Charles M. Voyton 1 , Meredith T. Morris , P. Christine Ackroyd 1 , James C. Morris , Kenneth A. Christensen 1
ACS Infectious Diseases ( IF 4.0 ) Pub Date : 2018-05-09 00:00:00 , DOI: 10.1021/acsinfecdis.8b00058 Charles M. Voyton 1 , Meredith T. Morris , P. Christine Ackroyd 1 , James C. Morris , Kenneth A. Christensen 1
Affiliation
|
Trypanosoma brucei, which causes human African typanosomiasis (HAT), derives cellular ATP from glucose metabolism while in the mammalian host. Targeting glucose uptake or regulation in the parasite has been proposed as a potential therapeutic strategy. However, few methods have been described to identify and characterize potential inhibitors of glucose uptake and regulation. Here, we report development of a screening assay that identifies small molecule disrupters of glucose levels in the cytosol and glycosomes. Using an endogenously expressed fluorescent protein glucose sensor expressed in cytosol or glycosomes, we monitored intracellular glucose depletion in the different cellular compartments. Two glucose level disrupters were identified, one of which only exhibited inhibition of glycosomal glucose and did not affect cytosolic levels. In addition to inhibiting glucose uptake with relatively high potency (EC50 = 700 nM), the compound also showed modest bloodstream form parasite killing activity. Expanding this assay will allow for identification of candidate compounds that disrupt parasite glucose metabolism.
中文翻译：
基于FRET流式细胞术的高通量筛选分析可鉴定布鲁氏锥虫中葡萄糖水平的破坏者
布氏锥虫会导致人类非洲肺吸虫病（HAT），在哺乳动物宿主体内时，其葡萄糖代谢会产生细胞ATP。已经提出了靶向寄生虫中葡萄糖的摄取或调节作为潜在的治疗策略。但是，很少有方法描述来鉴定和表征潜在的葡萄糖摄取和调节抑制剂。在这里，我们报告了一种筛选测定法的发展，该测定法可确定胞浆和糖体中葡萄糖水平的小分子干扰物。使用在细胞溶质或糖体中表达的内源性表达的荧光蛋白葡萄糖传感器，我们监测了不同细胞区室中的细胞内葡萄糖消耗。鉴定出两种葡萄糖水平破坏剂，其中一种仅表现出对糖体葡萄糖的抑制作用，并且不影响细胞溶质水平。50 = 700 nM），该化合物还显示出适度的血液形式的寄生物杀伤活性。扩展该测定将允许鉴定破坏寄生虫葡萄糖代谢的候选化合物。
更新日期：2018-05-09
中文翻译：
基于FRET流式细胞术的高通量筛选分析可鉴定布鲁氏锥虫中葡萄糖水平的破坏者
布氏锥虫会导致人类非洲肺吸虫病（HAT），在哺乳动物宿主体内时，其葡萄糖代谢会产生细胞ATP。已经提出了靶向寄生虫中葡萄糖的摄取或调节作为潜在的治疗策略。但是，很少有方法描述来鉴定和表征潜在的葡萄糖摄取和调节抑制剂。在这里，我们报告了一种筛选测定法的发展，该测定法可确定胞浆和糖体中葡萄糖水平的小分子干扰物。使用在细胞溶质或糖体中表达的内源性表达的荧光蛋白葡萄糖传感器，我们监测了不同细胞区室中的细胞内葡萄糖消耗。鉴定出两种葡萄糖水平破坏剂，其中一种仅表现出对糖体葡萄糖的抑制作用，并且不影响细胞溶质水平。50 = 700 nM），该化合物还显示出适度的血液形式的寄生物杀伤活性。扩展该测定将允许鉴定破坏寄生虫葡萄糖代谢的候选化合物。
京公网安备 11010802027423号