A rapid library screen for tailoring β-peptide structure and function
A rapid library screen for tailoring β-peptide structure and function
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DOI：
10.1021/ja055050o
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发表时间：
2005-10-26
影响因子：
15
通讯作者：
Schepartz, A
中科院分区：
文献类型：
--
作者：
Kritzer, JA;Luedtke, NW;Schepartz, A
最近，我们描述了一种β-十肽（β53-1），它在水溶液中折叠成14螺旋，以亚微摩尔亲和力结合癌蛋白hDM2，并抑制hDM2与p53激活域衍生的肽（p53AD）的相互作用。CD3OH中β53-1的溶液结构揭示了一个意想不到的c端解绕，它错开了包含hDM2识别表位的侧链，以更好地模仿p53AD。这种扭曲所隐含的结构-功能关系表明，沿非识别面具有多样性的β53-1类似物库可能包含对hDM2具有更大亲和力的分子。在这里，我们描述了(1)β-肽合成方案，产生高质量的一粒-一粒β-肽文库，适用于无需纯化的头上筛选，(2)一个通用的，可扩展的头上筛选，以及(3)一个简单的串联质谱（MS/MS）解码方法。利用这种方法，我们鉴定出具有改进的结构和功能特性的β53-1类似物。
Recently we described a β-decapeptide (β53-1) that folds into a 14-helix in aqueous solution, binds the oncoprotein hDM2 with submicromolar affinity, and inhibits the interaction of hDM2 with a peptide derived from the activation domain of p53 (p53AD). The solution structure ofβ53-1in CD3OH revealed an unexpected C-terminal unwinding that staggers the side chains comprising the hDM2 recognition epitope to better mimic those of p53AD. The structure−function relationship implied by this distortion suggested that a library ofβ53-1analogues possessing diversity along a nonrecognition face might contain molecules possessing greater affinity for hDM2. Here we describe (1) β-peptide synthesis protocols that produce high quality one-bead-one-β-peptide libraries suitable for on-bead screening without purification, (2) a versatile, scalable on-bead screen, and (3) a simple tandem mass spectrometry (MS/MS) decoding method. Using this procedure, we identifiedβ53-1analogues with improved structural and functional properties.