Activation of Cell-Penetrating Peptides with lonpair-π Interactions and Fluorophiles
Activation of Cell-Penetrating Peptides with lonpair-π Interactions and Fluorophiles
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DOI：
10.1021/jacs.6b06253
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发表时间：
2016-09-07
影响因子：
15
通讯作者：
Matile, Stefan
中科院分区：
文献类型：
--
作者：
Chuard, Nicolas;Fujisawa, Kaori;Matile, Stefan
在本报告中，我们阐述了激活富含精氨酸的细胞穿透肽（CPPs）的两个新概念。早期，我们认为，斥力驱动的离子配对相互作用与阴离子脂质解释了它们跨越疏水细胞膜的能力，疏水阴离子如芘丁酸盐可以加速这一过程，在动力学上胜过内体捕获。原来的解释是，苯乙烯丁酸酯的高活性可能源于CPP和活化剂之间的离子对- π相互作用，这意味着用极化推拉芳烃取代pi碱性芘可以获得更强的CPP活化剂。为了详细说明这一假设，我们准备了一小部分阴离子两亲体，它们可以通过离子对- π相互作用识别阳离子。与理论预测一致，我们发现平行而非反平行的离子对- π相互作用在模型膜和细胞中提供可操作的CPP激活剂。用全氟脂肪酸探讨了芘丁酸酯的高活性可能源于膜内自组装的另一种说法。它们的亲氟性有望促进膜中的自组装，而它们的高酸度应该阻止自组装反应中的电荷中和，即产生排斥驱动的离子配对相互作用。与这些预期一致，我们发现全氟脂肪酸在HeLa细胞中是强大的CPP激活剂，但在模型膜中不是。这些发现支持平行离子对- π相互作用和自组装亲氟试剂的排斥驱动离子配对作为激活CPPs的创新概念。这些结果也为反介导摄取作为富含精氨酸的cps的生产方式提供了更多的确证支持。
In this report, we elaborate on two new concepts to activate arginine-rich cell-penetrating peptides (CPPs). Early on, we have argued that repulsion-driven ion pairing interactions with anionic lipids account for their ability to move across hydrophobic cell membranes and that hydrophobic anions such as pyrenebutyrate can accelerate this process to kinetically outcompete endosomal capture. The original explanation that the high activity of pyrenebutyrate might originate from ionpair-pi interactions between CPP and activator implied that replacement of the pi-basic pyrene with polarized push-pull aromatics should afford more powerful CPP activators. To elaborate on this hypothesis, we prepared a small collection of anionic amphiphiles that could recognize cations by ionpair-pi interactions. Consistent with theoretical predictions, we find that parallel but not antiparallel ionpair-pi interactions afford operational CPP activators in model membranes and cells. The alternative suggestion that the high activity of pyrenebutyrate might originate from self-assembly in membranes was explored with perfluorinated fatty acids. Their fluorophilicity was expected to promote self-assembly in membranes, while their high acidity should prevent charge neutralization in response to self-assembly, i.e., generate repulsion-driven ion-pairing interactions. Consistent with these expectations, we find that perfluorinated fatty acids are powerful CPP activators in HeLa cells but not in model membranes. These findings support parallel ionpair-pi interactions and repulsion-driven ion pairing with self-assembled fluorophiles as innovative concepts to activate CPPs. These results also add much corroborative support for counterion-mediated uptake as the productive mode of action of arginine-rich CPPs.