A two-component 'double-click' approach to peptide stapling
A two-component 'double-click' approach to peptide stapling
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DOI：
10.1038/nprot.2015.033
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发表时间：
2015-04-01
期刊：
影响因子：
14.8
通讯作者：
Spring, David R.
中科院分区：
文献类型：
--
作者：
Lau, Yu Heng;Wu, Yuteng;Spring, David R.
多肽环化是一种有用的策略，可以将短的柔性多肽稳定成明确的生物活性构象，从而增强它们与蛋白质和其他重要生物分子相互作用的能力。在铜(I)催化下，我们提出了一种优化的方法来稳定具有α-螺旋构象的线形二叠氮多肽与二芳基连接物反应。由于这个过程产生带有双三唑键的侧链环化肽，所以它被称为‘双点击’装订。双击装订可以提高多肽抑制剂的结合亲和力、蛋白降解稳定性和细胞活性。双键装订的一个显著特点是，可以高效地合成具有不同主链连接的多肽，从而允许对多肽生物活性进行模块化控制。该方案描述了1，3-二烷基苯连接体和含有叠氮鸟氨酸的多肽之间的双击反应。还描述了后续的多肽纯化和确认步骤。整个双击装订方案可以在类似于48小时的时间内完成，包括两个隔夜的冷冻干燥步骤。
Peptide cyclization is a useful strategy for the stabilization of short flexible peptides into well-defined bioactive conformations, thereby enhancing their ability to interact with proteins and other important biomolecules. We present an optimized procedure for the stabilization of linear diazido peptides in an alpha-helical conformation upon reaction with dialkynyl linkers under Cu(I) catalysis. As this procedure generates side chain-cyclized peptides bearing a bis-triazole linkage, it is referred to as 'double-click' stapling. Double-click stapling can enhance the binding affinity, proteolytic stability and cellular activity of a peptide inhibitor. A distinguishing feature of double-click stapling is the efficiency with which peptides bearing different staple linkages can be synthesized, thus allowing for modular control over peptide bioactivity. This protocol describes the double-click reaction between a 1,3-dialkynylbenzene linker and peptides that contain azidoornithine. Subsequent peptide purification and confirmation steps are also described. The entire double-click stapling protocol can be completed in similar to 48 h, including two overnight lyophilization steps.