Kinetic analysis of cellular internalization and expulsion of unstructured D‐chirality cell penetrating peptides
Kinetic analysis of cellular internalization and expulsion of unstructured D‐chirality cell penetrating peptides
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DOI：
10.1002/aic.17087
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发表时间：
2020-10
期刊：
影响因子：
3.7
通讯作者：
Manibarathi Vaithiyanathan;Hannah C. Hymel;N. Safa;O. Sanchez;Jacob Pettigrew;C. Kirkpatrick;T. Gauthier;Adam T. Melvin
中科院分区：
文献类型：
--
作者：
Manibarathi Vaithiyanathan;Hannah C. Hymel;N. Safa;O. Sanchez;Jacob Pettigrew;C. Kirkpatrick;T. Gauthier;Adam T. Melvin
大多数细胞穿透肽（CPP）是非结构化的，易受蛋白水解降解。一种替代方案是将D-手性氨基酸掺入非结构化CPP中，以增强摄取和细胞内稳定性。这项工作使用一系列时间，浓度，温度和能量依赖性研究来研究CPP内化，导致D-手性肽的摄取增加3倍，稳定性增加50倍。CPP内化通过直接渗透和内吞作用的组合发生，其中一定百分比的内化CPP以时间依赖性方式从细胞中排出。机制研究发现，细胞通过胞吐作用独立途径输出完整的内化D-手性CPP，类似于直接渗透方法。这些发现突出了D手性CPP作为生物载体在治疗和生物传感应用中的潜力，但也确定了一种新的排出方法，表明摄取动力学，细胞内稳定性和输出动力学之间的关系。
Most cell penetrating peptides (CPPs) are unstructured and susceptible to proteolytic degradation. One alternative is to incorporate D‐chirality amino acids into unstructured CPPs to allow for enhanced uptake and intracellular stability. This work investigates CPP internalization using a series of time, concentration, temperature, and energy dependent studies, resulting in a three‐fold increase in uptake and 50‐fold increase in stability of D‐chirality peptides over L‐chirality counterparts. CPP internalization occurred via a combination of direct penetration and endocytosis, with a percentage of internalized CPP expelling from cells in a time‐dependent manner. Mechanistic studies identified that cells exported the intact internalized D‐chirality CPPs via an exocytosis independent pathway, analogous to a direct penetration method out of the cells. These findings highlight the potential of a D‐chirality CPP as bio‐vector in therapeutic and biosensing applications, but also identify a new expulsion method suggesting a relationship between uptake kinetics, intracellular stability, and export kinetics.