Bestatin-based chemical biology strategy reveals distinct roles for malaria M1-and M17-family aminopeptidases
Bestatin-based chemical biology strategy reveals distinct roles for malaria M1-and M17-family aminopeptidases
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DOI：
10.1073/pnas.1105601108
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发表时间：
2011-08-23
影响因子：
11.1
通讯作者：
Greenbaum, Doron C.
中科院分区：
文献类型：
--
作者：
Harbut, Michael B.;Velmourougane, Geetha;Greenbaum, Doron C.
疟疾导致世界范围内的发病率和死亡率，虽然化疗仍然是控制疟疾的极好手段，但耐药寄生虫需要发现新的抗疟药。肽酶是一类有前途的药物靶点，在恶性疟原虫红细胞生命周期中发挥着多种重要作用。在此，我们报告了一项多学科努力，结合基于活性的蛋白质分析、生化和肽组学方法，对两种遗传必需的恶性疟原虫金属氨基肽酶（MAP）——PfA-M1和Pf-LAP进行功能分析。通过合成一套基于通用 MAP 抑制剂支架 bestatin 的基于活性的探针 (ABP)，我们生成了这两种酶的特异性 ABP。 PfA-M1 的特异性抑制会导致寄生虫消化液泡肿胀，并阻止血红蛋白 (Hb) 衍生的寡肽的蛋白水解，可能使寄生虫挨饿导致死亡。相反，在 Hb 降解开始之前，Pf-LAP 的抑制在生命周期的早期对寄生虫是致命的，这表明 Pf-LAP 在 Hb 消化之外具有重要作用。
Malaria causes worldwide morbidity and mortality, and while chemotherapy remains an excellent means of malaria control, drug-resistant parasites necessitate the discovery of new antimalarials. Peptidases are a promising class of drug targets and perform several important roles during the Plasmodium falciparum erythrocytic life cycle. Herein, we report a multidisciplinary effort combining activity-based protein profiling, biochemical, and peptidomic approaches to functionally analyze two genetically essential P. falciparum metallo-aminopeptidases (MAPs), PfA-M1 and Pf-LAP. Through the synthesis of a suite of activity-based probes (ABPs) based on the general MAP inhibitor scaffold, bestatin, we generated specific ABPs for these two enzymes. Specific inhibition of PfA-M1 caused swelling of the parasite digestive vacuole and prevented proteolysis of hemoglobin (Hb)-derived oligopeptides, likely starving the parasite resulting in death. In contrast, inhibition of Pf-LAP was lethal to parasites early in the life cycle, prior to the onset of Hb degradation suggesting that Pf-LAP has an essential role outside of Hb digestion.