Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne , CH-1015 Lausanne, Switzerland.
J Am Chem Soc. 2014 Apr 23;136(16):5880-3. doi: 10.1021/ja501861m. Epub 2014 Apr 11.
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.
光致变色配体是一种强大的工具，可以在高时空分辨率下控制生物过程。不幸的是，这样的配体只存在于有限数量的蛋白质中，而且通过合理设计来开发它们并不简单。我们开发了一种体外进化策略，以生成针对所选靶标的光激活肽配体。简而言之，随机肽通过噬菌体展示进行编码，用偶氮苯接头进行化学环化，用紫外线照射将偶氮苯转化为顺式构象，然后针对模型靶标链霉亲和素进行淘选。分离出的肽具有强烈的共有序列，表明具有靶标特异性结合。当与偶氮苯接头环化时，一些肽具有很高的亲和力，并且它们的亲和力可以通过紫外线调节。所提出的方法具有鲁棒性，可以应用于几乎任何目标的光致变色配体的体外进化。
