我们以往的工作发现在所筛选的一系列G蛋白偶联受体中，只有大鼠胰腺腺泡细胞所表达的CCK1型受体，具有可被光动力作用所产生的单线态氧分子所永久激活的“嘎嘣脆”特性。本项目据此研究在新鲜分离的其它类型细胞包括中枢和外周神经元、外周器官细胞的CCK1型受体，以及在不同种类细胞系所异源表达的CCK1型受体，是否同样具备被光动力永久氧化激活的“嘎嘣脆”特性。将使用基因编码的蛋白质光敏剂miniSOG或miniSOG-CCK1R融合蛋白，获得光动力永久激活CCK1型受体的最佳条件；包括通过NanoLuc-miniSOG生物发光永久性光动力激活CCK1R受体。尝试将CCK1型受体的“嘎嘣脆”特性引入其它G蛋白偶联受体，光动力永久激活CCK1R-XnR嵌合体受体。研究CCK1型受体被光动力永久激活或“嘎嘣脆”特性的进化历程，为将“嘎嘣脆”受体用于在体动物试验奠定细胞生理学基础。

We have previously found that among various G protein-coupled receptors screened, only pancreatic acinar cell CCK1 could be permanently activated by photodynamically-generated singlet oxygen, we therefore named CCK1 receptor a G Protein-Coupled Receptor Activated By Singlet Oxygen (GPCR-ABSO). In the present project we will examine whether CCK1 receptor expressed in other cell types both central and peripheral, and CCK1 receptor ectopically expressed in different cell lines are also permanently activated photodynamically. Photodynamic activation of CCK1 receptor with genetically encoded protein photosensitisers such as miniSOG will be optimized, and the possibility of using light from NanoLuc bioluminescence to power the miniSOG photodynamic activation of CCK1 receptors will be examined. We will also attempt to introduce the GPCR-ABSO property to other G protein-coupled receptors to generate chimeric receptors CCK1R-XnR. Evolution of CCK1 receptor photodynamic activation will also be examined. These works will provide the cellular basis for in vivo application of GPCR-pABSO in the near future.