Over two decades, γ-secretase has been the target for extensive therapeutic development due to its pivotal role in pathogenesis of Alzheimer’s disease and cancer. However, it has proven to be a challenging task owing to its large set of substrates and our limited understanding of the enzyme’s structural and mechanistic features. The scientific community is taking bigger strides towards solving this puzzle with recent advancement in techniques like cryogenic electron microscopy (cryo-EM) and photo-affinity labelling (PAL). This review highlights the significance of the PAL technique with multiple examples of photo-probes developed from γ-secretase inhibitors and modulators. The binding of these probes into active and/or allosteric sites of the enzyme has provided crucial information on the γ-secretase complex and improved our mechanistic understanding of this protease. Combining the knowledge of function and regulation of γ-secretase will be a decisive factor in developing novel γ-secretase modulators and biological therapeutics.

二十多年来，由于γ-分泌酶在阿尔茨海默氏病和癌症的发病机理中起着关键作用，因此一直是广泛治疗发展的目标。然而，由于其底物种类繁多以及我们对酶的结构和机理特征的了解有限，因此已被证明是一项艰巨的任务。随着低温电子显微镜（cryo-EM）和光亲和标记（PAL）等技术的最新发展，科学界正在朝着解决这一难题的方向迈出更大的步伐。这篇综述突出了PAL技术的重要性，其中列举了多个由γ-分泌酶抑制剂和调节剂开发的光探针实例。这些探针与酶的活性和/或变构位点的结合为γ-分泌酶复合物提供了关键信息，并改善了我们对该蛋白酶的机械理解。结合γ-分泌酶的功能和调节知识将是开发新型γ-分泌酶调节剂和生物疗法的决定性因素。