Comparative chemical genomics reveal that the spiroindolone antimalarial KAE609 (Cipargamin) is a P-type ATPase inhibitor.
Comparative chemical genomics reveal that the spiroindolone antimalarial KAE609 (Cipargamin) is a P-type ATPase inhibitor.
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DOI：
10.1038/srep27806
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发表时间：
2016-06-13
影响因子：
4.6
通讯作者：
Winzeler EA
中科院分区：
文献类型：
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作者：
Goldgof GM;Durrant JD;Ottilie S;Vigil E;Allen KE;Gunawan F;Kostylev M;Henderson KA;Yang J;Schenken J;LaMonte GM;Manary MJ;Murao A;Nachon M;Murray R;Prescott M;McNamara CW;Slayman CW;Amaro RE;Suzuki Y;Winzeler EA
螺隆酮是在细胞筛选中发现的一类新的抗疟疾药物，它由于寄生虫p型atp酶PfATP4的突变而活性降低。我们在这里表明，酿酒葡萄球菌在暴露于螺隆酮后也获得编码p型atp酶（ScPMA1）的基因突变，这些突变足以产生耐药性。ScPMA1的KAE609耐药突变不赋予对不相关抗菌素的抗性，但赋予对烷基溶血磷脂雪草碱的交叉敏感性，已知其将ScPMA1从质膜上取代。通过体外无细胞实验，我们证明KAE609直接抑制ScPma1p atp酶活性。KAE609还能增加酵母细胞的胞质氢离子浓度。计算机对接到ScPma1p同源模型，确定了一种结合模式，支持恶性疟原虫和酿酒葡萄球菌的遗传抗性决定因素和体外实验结构-活性关系。该模型还提示与二氢异喹诺酮类抗疟药物有一个共同的结合位点。我们的数据支持KAE609通过直接干扰p型atp酶活性发挥其抗疟疾活性的模型。
The spiroindolones, a new class of antimalarial medicines discovered in a cellular screen, are rendered less active by mutations in a parasite P-type ATPase, PfATP4. We show here that S. cerevisiae also acquires mutations in a gene encoding a P-type ATPase (ScPMA1) after exposure to spiroindolones and that these mutations are sufficient for resistance. KAE609 resistance mutations in ScPMA1 do not confer resistance to unrelated antimicrobials, but do confer cross sensitivity to the alkyl-lysophospholipid edelfosine, which is known to displace ScPma1p from the plasma membrane. Using an in vitro cell-free assay, we demonstrate that KAE609 directly inhibits ScPma1p ATPase activity. KAE609 also increases cytoplasmic hydrogen ion concentrations in yeast cells. Computer docking into a ScPma1p homology model identifies a binding mode that supports genetic resistance determinants and in vitro experimental structure-activity relationships in both P. falciparum and S. cerevisiae. This model also suggests a shared binding site with the dihydroisoquinolones antimalarials. Our data support a model in which KAE609 exerts its antimalarial activity by directly interfering with P-type ATPase activity.