Phage Selection of Photoswitchable Peptide Ligands
Phage Selection of Photoswitchable Peptide Ligands
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DOI：
10.1021/ja501861m
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发表时间：
2014-04-23
影响因子：
15
通讯作者：
Heinis, Christian
中科院分区：
文献类型：
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作者：
Bellotto, Silvia;Chen, Shiyu;Heinis, Christian
光开关配体是在高空间和时间分辨率下控制生物过程的有力工具。不幸的是，这样的配体只存在于有限数量的蛋白质中，它们通过合理设计的发展不是微不足道的。我们已经开发了一种体外进化策略，以产生光激活肽配体的选择目标。简而言之，随机肽通过噬菌体展示编码，用偶氮苯连接物进行化学环化，暴露在紫外线下将偶氮苯转换成顺式构象，并与模型目标链霉亲和素进行平移。分离的肽具有很强的一致性序列，表明特异性结合。一些肽与偶氮苯连接环结合时具有高亲和力，并且它们的亲和力可以通过紫外线调节。所提出的方法是稳健的，可以应用于光开关配体的体外进化到几乎任何目标。
Photoswitchable ligands are powerful tools to control biological processes at high spatial and temporal resolution. Unfortunately, such ligands exist only for a limited number of proteins and their development by rational design is not trivial. We have developed an in vitro evolution strategy to generate light-activatable peptide ligands to targets of choice. In brief, random peptides were encoded by phage display, chemically cyclized with an azobenzene linker, exposed to UV light to switch the azobenzene into cis conformation, and panned against the model target streptavidin. Isolated peptides shared strong consensus sequences, indicating target-specific binding. Several peptides bound with high affinity when cyclized with the azobenzene linker, and their affinity could be modulated by UV light. The presented method is robust and can be applied for the in vitro evolution of photoswitchable ligands to virtually any target.