SARS-CoV-2 NSP2 as a Potential Delivery Vehicle for Proteins
SARS-CoV-2 NSP2 as a Potential Delivery Vehicle for Proteins
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DOI：
10.1021/acs.molpharmaceut.3c00680
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发表时间：
2024-01-30
影响因子：
4.9
通讯作者：
Zhang，Guigen
中科院分区：
文献类型：
--
作者：
Zheng，Ningze;Liu，Shurui;Zhang，Guigen
生物分子递送系统的开发对于治疗各种疾病如癌症、免疫性疾病和代谢紊乱是必不可少的。我们首次发现SARS-CoV-2编码的非结构蛋白2（NSP 2）可以从细胞中分泌出来，并在细胞中合成。ER/高尔基体分泌途径的抑制剂Brefeldin A和H89不抑制NSP 2的分泌。NSP 2可能通过非常规分泌途径分泌。此外，分泌的和纯化的NSP 2蛋白都能够穿过质膜屏障并进入永生化的人脐静脉内皮细胞和肿瘤细胞系。进入后，NSP 2蛋白仅定位于细胞质中。细胞松弛素D是一种有效的肌动蛋白聚合抑制剂，可抑制NSP 2的进入。NSP 2可以携带其他分子进入细胞。伯克霍尔德氏菌致死因子1是一种来自胞内病原体类鼻疽伯克霍尔德氏菌的单体毒素，已证明其通过靶向宿主真核起始翻译因子4A而具有抗肿瘤活性。NSP 2-BLF 1融合蛋白跨Huh 7细胞的细胞膜易位并介导细胞杀伤。通过使用不同的方法，包括蛋白质纯化，化学抑制和细胞成像，我们证实NSP 2能够将异源蛋白质递送到细胞中。NSP 2可以作为蛋白质的潜在递送载体。
The development of biomolecule delivery systems is essential for the treatment of various diseases such as cancer, immunological diseases, and metabolic disorders. For the first time, we found that SARS-CoV-2-encoded nonstructural protein 2 (NSP2) can be secreted from the cells, where it is synthesized. Brefeldin A and H89, inhibitors of ER/Golgi secretion pathways, did not inhibit NSP2 secretion. NSP2 is likely secreted via an unconventional secretory pathway. Moreover, both secreted and purified NSP2 proteins were able to traverse the plasma membrane barrier and enter both immortalized human umbilical vein endothelial cells and tumor cell lines. After entry, the NSP2 protein was localized in only the cytoplasm. Cytochalasin D, a potent inhibitor of actin polymerization, inhibited the entry of NSP2. NSP2 can carry other molecules into cells. Burkholderia lethal factor 1, a monomeric toxin from the intracellular pathogenBurkholderia pseudomallei, has demonstrated antitumor activity by targeting host eukaryotic initiation translation factor 4A. An NSP2-BLF1 fusion protein was translocated across the cellular membranes of Huh7 cells and mediated cell killing. By using different approaches, including protein purification, chemical inhibition, and cell imaging, we confirm that NSP2 is able to deliver heterologous proteins into cells. NSP2 can act as a potential delivery vehicle for proteins.