CXC-Mediated Cellular Uptake of Miniproteins: Forsaking “Arginine Magic”
CXC-Mediated Cellular Uptake of Miniproteins: Forsaking “Arginine Magic”
复制标题
CXC 介导的细胞对微量蛋白的摄取：放弃“精氨酸魔法”
DOI：
10.1021/acschembio.8b00564
复制
发表时间：
2018
影响因子：
4
通讯作者：
Chuanliu Wu
中科院分区：
文献类型：
--
作者：
Xiaoting Meng;Tao Li;Yibing Zhao;Chuanliu Wu
微蛋白的大小介于较大的生物制剂和小分子之间，并且可能具有两者的优点;它们代表了用于设计亲和试剂、酶和治疗剂的一类有前途的支架。促进微蛋白的细胞摄取的常规策略依赖于精氨酸残基的广泛接枝或嵌入。然而，使用阳离子精氨酸的要求会给修饰的小蛋白带来问题，例如，在溶液中的低溶解度（聚集倾向）和潜在的毒性，这是肽和蛋白质社区中的公开秘密。在这项工作中，我们报告的阳离子miniproteins的细胞渗透性可以进一步显着增加，通过附加一个神奇的CXC（半胱氨酸-任何-半胱氨酸）基序，这利用了巯基-二硫键交换的细胞表面。更重要的是，我们发现当嵌入的精氨酸全部被赖氨酸残基取代时，附加CXC的小蛋白的高细胞渗透性仍然可以被保留，这表明几乎所有细胞渗透性肽和（小）蛋白所必需的“精氨酸魔术”对于CXC介导的细胞摄取是不需要的。这一发现提供了一种新的途径，用于设计高细胞渗透性的微蛋白，而不损害潜在的毒性和稳定性所产生的精氨酸嵌入或接枝。
Miniproteins have a size between that of larger biologics and small molecules and presumably possess the advantages of both; they represent an expanding class of promising scaffolds for the design of affinity reagents, enzymes, and therapeutics. Conventional strategies to promote cellular uptake of miniproteins rely on extensive grafting or embedding of arginine residues. However, the requirement of using cationic arginines would cause problems to the modified miniproteins, for example, low solubility in solutions (proneness of aggregation) and potential toxicity, which are open secrets in the peptide and protein communities. In this work, we report that the cell-permeability of cationic miniproteins can be further markedly increased through appending a magic CXC (cysteine-any-cysteine) motif, which takes advantage of thiol–disulfide exchanges on the cell surface. More importantly, we discovered that the high cell permeability of the CXC-appended miniproteins can still be preserved when the embedded arginines are all substituted with lysine residues, indicating that the “arginine magic” essential to almost all cell-permeable peptides and (mini)proteins is not required for the CXC-mediated cellular uptake. This finding provides a new avenue for designing highly cell-permeable miniproteins without compromise of potential toxicity and stability arising from arginine embedding or grafting.