Boucher Lauren E, Hopp Christine S, Muthinja Julianne Mendi, Frischknecht Friedrich, Bosch Jürgen
Department of Biochemistry and Molecular Biology , Johns Hopkins Bloomberg School of Public Health , 615 N. Wolfe Street , Baltimore , Maryland 21205 , United States.
Johns Hopkins Malaria Research Institute , Johns Hopkins Bloomberg School of Public Health , 615 N. Wolfe Street , Baltimore , Maryland 21205 , United States.
ACS Infect Dis. 2018 Apr 13;4(4):620-634. doi: 10.1021/acsinfecdis.7b00225. Epub 2018 Feb 19.
As obligate, intracellular parasites, Plasmodium spp. rely on invasion of host cells in order to replicate and continue their life cycle. The parasite needs to traverse the dermis and endothelium of blood vessels, invade hepatocytes and red blood cells, traverse the mosquito midgut, and enter the salivary glands to continue the cycle of infection and transmission. To traverse and invade cells, the parasite employs an actomyosin motor at the core of a larger invasion machinery complex known as the glideosome. The complex is comprised of multiple protein-protein interactions linking the motor to the internal cytoskeletal network of the parasite and to the extracellular adhesins, which directly contact the host tissue or cell surface. One key interaction is between the cytoplasmic tails of the thrombospondin related anonymous protein (TRAP) and aldolase, a bridging protein to the motor. Here, we present results from screening the Medicines for Malaria Venture (MMV) library of 400 compounds against this key protein-protein interaction. Using a surface plasmon resonance screen, we have identified several compounds that modulate the dynamics of the interaction between TRAP and aldolase. These compounds have been validated in vitro by studying their effects on sporozoite gliding motility and hepatocyte invasion. One of the MMV compounds identified reduced invasion levels by 89% at the lowest concentration tested (16 μM) and severely inhibited gliding at even lower concentrations (5 μM). By targeting protein-protein interactions, we investigated an under-explored area of parasite biology and general drug development, to identify potential antimalarial lead compounds.
作为专性细胞内寄生虫，疟原虫属依赖侵入宿主细胞来进行复制并延续其生命周期。该寄生虫需要穿过血管的真皮和内皮，侵入肝细胞和红细胞，穿过蚊子的中肠，并进入唾液腺以延续感染和传播循环。为了穿过并侵入细胞，寄生虫在一种称为滑行体的更大的侵入机制复合体的核心部位利用了一种肌动球蛋白马达。该复合体由多种蛋白质 - 蛋白质相互作用组成，这些相互作用将马达与寄生虫的内部细胞骨架网络以及与直接接触宿主组织或细胞表面的细胞外粘附素相连。一种关键的相互作用发生在血小板反应蛋白相关无名蛋白（TRAP）的细胞质尾巴与醛缩酶之间，醛缩酶是连接到马达的一种桥梁蛋白。在此，我们展示了针对这种关键蛋白质 - 蛋白质相互作用对疟疾药物事业（MMV）的400种化合物库进行筛选的结果。通过表面等离子体共振筛选，我们鉴定出了几种调节TRAP与醛缩酶之间相互作用动态的化合物。这些化合物已通过研究它们对子孢子滑行运动和肝细胞侵入的影响在体外得到验证。所鉴定出的一种MMV化合物在测试的最低浓度（16 μM）下使侵入水平降低了89%，并且在更低浓度（5 μM）时严重抑制了滑行。通过靶向蛋白质 - 蛋白质相互作用，我们研究了寄生虫生物学和一般药物开发中一个未充分探索的领域，以鉴定潜在的抗疟先导化合物。
