Schisandrin A is a natural dibenzocyclooctene lignan with potent neuroprotective activity. However, the specific mechanisms or direct target proteins have not been clarified up to now. In this study, we designed and synthesized the probes of schisandrin A with photoreactive diazirine and clickable alkyne to identify its direct target in SH-SY5Y cells by employing activity-based protein profiling (ABPP) technique. Ykt6 was prominent among the 13 proteins obtained with high confidence and we confirmed Ykt6 as the direct target of schisandrin A by CETSA, IF, SPR and knockdown assay. Functionally, schisandrin A protected the cells against the injury induced by glutamate by regulating autophagy via Ykt6. This discovery may provide a novel therapeutic option for various neuronal cell damage-mediated diseases.

Schisandrin A 是一种天然的二苯并环辛烯木酚素，具有强大的神经保护活性。然而，具体机制或直接靶蛋白迄今尚未阐明。在这项研究中，我们设计并合成了五味子甲素与光反应性二氮丙啶和可点击炔烃的探针，通过采用基于活性的蛋白质谱分析 (ABPP) 技术鉴定其在 SH-SY5Y 细胞中的直接靶标。Ykt6 在以高置信度获得的 13 种蛋白质中最为突出，我们通过 CETSA、IF、SPR 和敲低测定证实 Ykt6 是五味子甲素的直接靶标。在功能上，五味子甲通过调节自噬来保护细胞免受谷氨酸诱导的损伤Ykt6。这一发现可能为各种神经元细胞损伤介导的疾病提供一种新的治疗选择。