Targeting a dynamic protein-protein interaction: fragment screening against the malaria myosin A motor complex.
Targeting a dynamic protein-protein interaction: fragment screening against the malaria myosin A motor complex.
复制标题
DOI：
10.1002/cmdc.201402357
复制
发表时间：
2015-01
期刊：
影响因子：
3.4
通讯作者：
Tate EW
中科院分区：
文献类型：
--
作者：
Douse CH;Vrielink N;Wenlin Z;Cota E;Tate EW
蠕动是恶性疟原虫复杂生命周期的一个重要特征，恶性疟原虫是导致人类疟疾的顶复门寄生虫。宿主细胞入侵等过程被认为是由保守的肌动球蛋白马达（包含肌球蛋白 A 或 myoA）提供动力，其正确定位取决于与寄生虫内膜的肌球蛋白 A 尾部结构域相互作用蛋白 (MTIP) 的紧密相互作用。尽管破坏这种蛋白质-蛋白质相互作用是研究基于 myoA 的运动和抑制寄生生命周期的假定作用的一种有吸引力的方法，但尚未发现与 MTIP 结合的小分子。此外，还不可能获得游离蛋白的晶体结构，该蛋白在缺乏天然 myoA 尾部伙伴的情况下是高度动态且不稳定的。在此，我们报告了通过基于片段的综合生物物理方法和结构研究，从头鉴定了第一个结合并稳定 MTIP 的分子，以检查命中化合物的结合模式。整个过程中都解决了使用传统片段筛选工作流程针对此类动态系统的挑战。
Motility is a vital feature of the complex life cycle of Plasmodium falciparum, the apicomplexan parasite that causes human malaria. Processes such as host cell invasion are thought to be powered by a conserved actomyosin motor (containing myosin A or myoA), correct localization of which is dependent on a tight interaction with myosin A tail domain interacting protein (MTIP) at the inner membrane of the parasite. Although disruption of this protein–protein interaction represents an attractive means to investigate the putative roles of myoA-based motility and to inhibit the parasitic life cycle, no small molecules have been identified that bind to MTIP. Furthermore, it has not been possible to obtain a crystal structure of the free protein, which is highly dynamic and unstable in the absence of its natural myoA tail partner. Herein we report the de novo identification of the first molecules that bind to and stabilize MTIP via a fragment-based, integrated biophysical approach and structural investigations to examine the binding modes of hit compounds. The challenges of targeting such a dynamic system with traditional fragment screening workflows are addressed throughout.