N-terminal signal peptides facilitate the engineering of PVC complex as a potent protein delivery system.
N-terminal signal peptides facilitate the engineering of PVC complex as a potent protein delivery system.
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N 端信号肽有助于将 PVC 复合物工程化为有效的蛋白质递送系统
DOI：
10.1126/sciadv.abm2343
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发表时间：
2022-04-29
期刊：
影响因子：
13.6
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中科院分区：
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细胞外收缩注射系统（eCIS）是广泛存在的类似于T4噬菌体尾的细菌纳米机器。光杆状病毒毒力盒（Photorhabdus virtual cassette，PVC）是一种典型的细胞外毒素注射系统。这使其成为生物医学研究中蛋白质递送的理想工具。然而，如何操纵这种纳米复合物作为一个分子注射器仍然没有确定。在这里，我们确定了一组N-末端信号肽（SP）序列是至关重要的效应加载到PVC复合物的内管。通过应用遗传操作、冷冻电镜、体外易位分析和动物实验，我们表明，在SP的指导下，许多原核和真核蛋白可以装载到PVC中，以发挥其跨细胞膜的功能。因此，我们可以定制PVC作为一个有效的蛋白质递送纳米注射器的生物治疗，通过选择货物蛋白在一个广泛的频谱，无论他们的物种，大小和电荷。
Extracellular contractile injection systems (eCISs) are widespread bacterial nanomachines that resemble T4 phage tail. As a typical eCIS, Photorhabdus virulence cassette (PVC) was proposed to inject toxins into eukaryotic cells by puncturing the cell membrane from outside. This makes it an ideal tool for protein delivery in biomedical research. However, how to manipulate this nanocomplex as a molecular syringe is still undetermined. Here, we identify that one group of N-terminal signal peptide (SP) sequences are crucial for the effector loading into the inner tube of PVC complex. By application of genetic operation, cryo–electron microscopy, in vitro translocation assays, and animal experiments, we show that, under the guidance of the SP, numerous prokaryotic and eukaryotic proteins can be loaded into PVC to exert their functions across cell membranes. We therefore might customize PVC as a potent protein delivery nanosyringe for biotherapy by selecting cargo proteins in a broad spectrum, regardless of their species, sizes, and charges.