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Designed Parasite-Selective Rhomboid Inhibitors Block Invasion and Clear Blood-Stage Malaria.
Cell Chemical Biology ( IF 8.6 ) Pub Date : 2020-09-03 , DOI: 10.1016/j.chembiol.2020.08.011
Shiv Gandhi 1 , Rosanna P Baker 1 , Sangwoo Cho 1 , Stancho Stanchev 2 , Kvido Strisovsky 2 , Siniša Urban 1
Affiliation  

Rhomboid intramembrane proteases regulate pathophysiological processes, but their targeting in a disease context has never been achieved. We decoded the atypical substrate specificity of malaria rhomboid PfROM4, but found, unexpectedly, that it results from “steric exclusion”: PfROM4 and canonical rhomboid proteases cannot cleave each other's substrates due to reciprocal juxtamembrane steric clashes. Instead, we engineered an optimal sequence that enhanced proteolysis >10-fold, and solved high-resolution structures to discover that boronates enhance inhibition >100-fold. A peptide boronate modeled on our “super-substrate” carrying one “steric-excluding” residue inhibited PfROM4 but not human rhomboid proteolysis. We further screened a library to discover an orthogonal alpha-ketoamide that potently inhibited PfROM4 but not human rhomboid proteolysis. Despite the membrane-immersed target and rapid invasion, ultrastructural analysis revealed that single-dosing blood-stage malaria cultures blocked host-cell invasion and cleared parasitemia. These observations establish a strategy for designing parasite-selective rhomboid inhibitors and expose a druggable dependence on rhomboid proteolysis in non-motile parasites.



中文翻译:

设计的寄生虫选择性菱形抑制剂阻止入侵和清除血期疟疾。

菱形膜内蛋白酶调节病理生理过程,但从未实现它们在疾病背景下的靶向。我们解码了疟疾菱形 PfROM4 的非典型底物特异性,但意外地发现它是由“空间排斥”引起的:由于相互的近膜空间冲突,PfROM4 和经典菱形蛋白酶不能切割彼此的底物。相反,我们设计了一个最佳序列,可将蛋白水解增强 > 10 倍,并解析高分辨率结构以发现硼酸盐可增强抑制 > 100 倍。以我们的“超级底物”为模型的肽硼酸盐携带一个“立体排除”残基抑制 PfROM4,但不抑制人菱形蛋白水解。我们进一步筛选了一个库,以发现有效抑制 PfROM4 但不抑制人类菱形蛋白水解的正交 α-酮酰胺。尽管膜浸入目标和快速侵袭,超微结构分析显示单剂量血阶段疟疾培养物阻止宿主细胞侵袭并清除寄生虫血症。这些观察结果建立了一种设计寄生虫选择性菱形抑制剂的策略,并揭示了非运动寄生虫对菱形蛋白水解的药物依赖性。

更新日期:2020-09-03
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