通过RaPID系统进行硫醚封闭大环肽配体的体外筛选。
In Vitro Selection of Thioether-Closed Macrocyclic Peptide Ligands by Means of the RaPID System.
作者信息
Katoh Takayuki, Goto Yuki, Suga Hiroaki
机构信息
Department of Chemistry, Graduate School of Science, The University of Tokyo, Tokyo, Japan.
出版信息
Methods Mol Biol. 2022;2371:247-259. doi: 10.1007/978-1-0716-1689-5_13.
The Random nonstandard Peptides Integrated Discovery (RaPID) system enables efficient screening of macrocyclic peptides with high affinities against target molecules. Random peptide libraries are prepared by in vitro translation using the Flexible In vitro Translation (FIT) system, which allows for incorporation of diverse nonproteinogenic amino acids into peptides by genetic code reprogramming. By introducing an N-chloroacetyl amino acid at the N-terminus and a Cys at the downstream, macrocyclic peptide libraries can be readily generated via posttranslational thioether formation. Here, we describe how to prepare a thioether-closed macrocyclic peptide library, and its application to the RaPID screening.
随机非标准肽整合发现（RaPID）系统能够高效筛选出对目标分子具有高亲和力的大环肽。使用灵活体外翻译（FIT）系统通过体外翻译制备随机肽库，该系统允许通过遗传密码重编程将多种非蛋白质ogenic氨基酸掺入肽中。通过在N端引入N-氯乙酰氨基酸并在下游引入半胱氨酸，可以通过翻译后硫醚形成轻松生成大环肽库。在此，我们描述了如何制备硫醚封闭的大环肽库及其在RaPID筛选中的应用。
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