Bastianelli Giacomo, Bouillon Anthony, Nguyen Christophe, Le-Nguyen Dung, Nilges Michael, Barale Jean-Christophe
Institut Pasteur, Unité de Bioinformatique Structurale, Département de Biologie Structurale et Chimie, Paris, France; CNRS UMR 3528, Paris, France.
Institut Pasteur, Unité d'Immunologie Moléculaires des Parasites, Département de Parasitologie et de Mycologie & CNRS URA 2581, Paris, France; CNRS, URA2581, Paris, France.
PLoS One. 2014 Oct 24;9(10):e109269. doi: 10.1371/journal.pone.0109269. eCollection 2014.
Malaria remains a major global health concern. The development of novel therapeutic strategies is critical to overcome the selection of multiresistant parasites. The subtilisin-like protease (SUB1) involved in the egress of daughter Plasmodium parasites from infected erythrocytes and in their subsequent invasion into fresh erythrocytes has emerged as an interesting new drug target.
Using a computational approach based on homology modeling, protein-protein docking and mutation scoring, we designed protein-based inhibitors of Plasmodium vivax SUB1 (PvSUB1) and experimentally evaluated their inhibitory activity. The small peptidic trypsin inhibitor EETI-II was used as scaffold. We mutated residues at specific positions (P4 and P1) and calculated the change in free-energy of binding with PvSUB1. In agreement with our predictions, we identified a mutant of EETI-II (EETI-II-P4LP1W) with a Ki in the medium micromolar range.
Despite the challenges related to the lack of an experimental structure of PvSUB1, the computational protocol we developed in this study led to the design of protein-based inhibitors of PvSUB1. The approach we describe in this paper, together with other examples, demonstrates the capabilities of computational procedures to accelerate and guide the design of novel proteins with interesting therapeutic applications.
疟疾仍然是全球主要的健康问题。开发新型治疗策略对于克服多重耐药寄生虫的选择至关重要。参与疟原虫子代从受感染红细胞中逸出并随后侵入新鲜红细胞的枯草杆菌蛋白酶样蛋白酶(SUB1)已成为一个有趣的新药物靶点。
我们基于同源建模、蛋白质-蛋白质对接和突变评分的计算方法,设计了间日疟原虫SUB1(PvSUB1)的蛋白质基抑制剂,并通过实验评估了它们的抑制活性。小肽胰蛋白酶抑制剂EETI-II用作支架。我们对特定位置(P4和P1)的残基进行了突变,并计算了与PvSUB1结合的自由能变化。与我们的预测一致,我们鉴定出一种EETI-II突变体(EETI-II-P4LP1W),其Ki在中微摩尔范围内。
尽管由于缺乏PvSUB1的实验结构而面临挑战,但我们在本研究中开发的计算方案导致了PvSUB1蛋白质基抑制剂的设计。我们在本文中描述的方法以及其他例子,证明了计算程序在加速和指导具有有趣治疗应用的新型蛋白质设计方面的能力。