Chemical synthesis of cell-permeable apoptotic peptides from in vivo produced proteins.
Chemical synthesis of cell-permeable apoptotic peptides from in vivo produced proteins.
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DOI：
10.1021/bc200338u
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发表时间：
2011-08
影响因子：
4.7
通讯作者：
T. Fricke;R. J. Mart;C. Watkins;M. Wiltshire;R. Errington;Paul J. Smith;A. Jones;R. Allemann
中科院分区：
文献类型：
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作者：
T. Fricke;R. J. Mart;C. Watkins;M. Wiltshire;R. Errington;Paul J. Smith;A. Jones;R. Allemann
细菌表达体内合成肽已发展成为固相法合成多肽的可靠替代品。体内方法的一个重大缺点是基因产物很难在翻译后进行修饰。在这里，我们提出了一种简单的方法来修饰细菌宿主中产生的多肽，这些多肽是在C-末端蛋氨酸上被溴化氰裂解后产生的。生成的高丝氨酸内酯与炔丙胺反应，可以用多种化学物质进行高效和选择性的修饰，如荧光染料、生物素衍生物、聚戊烯、脂类、多糖或多肽。细胞穿透性多肽8-精氨酸(R(8))与来自促凋亡肿瘤抑制因子Bak BH3的多肽结合，导致细胞有效摄取并随后从线粒体释放细胞色素c，最终诱导细胞凋亡，与通过肽骨架与R(8)相连的多肽相似。这些结果突出了在活细胞中使用这种工具的巨大潜力。
In vivo synthesis of peptides by bacterial expression has developed into a reliable alternative to solid-phase peptide synthesis. A significant drawback of in vivo methods is the difficulty with which gene products can be modified post-translationally. Here, we present a method for the facile modification of peptides generated in bacterial hosts after cyanogen bromide cleavage at C-terminal methionines. Reaction of the resulting homoserine lactones with propargylamine allows efficient and selective modification with a wide variety of chemicals such as fluorescent dyes, biotin derivatives, polyprenyls, lipids, polysaccharides, or peptides. Attachment of the cell penetrating peptide octa-arginine (R(8)) to peptides derived from the proapoptotic tumor suppressor Bak BH3 led to efficient cellular uptake and subsequent cytochrome c release from mitochondria, culminating in induction of apoptosis similar to that observed with peptides linked to R(8) via the peptide backbone. These results highlight the significant potential for use of such tools in live cells.