Liver Cancer Institute, Zhongshan Hospital and Department of Chemistry, Fudan University, Shanghai, China.
Institutes of Biomedical Sciences and NHC Key Laboratory of Glycoconjugates Research, Shanghai, China.
Nat Commun. 2024 Apr 8;15(1):2997. doi: 10.1038/s41467-024-47033-w.
Cell surface proteins are responsible for many crucial physiological roles, and they are also the major category of drug targets as the majority of therapeutics target membrane proteins on the surface of cells to alter cellular signaling. Despite its great significance, ligand discovery against membrane proteins has posed a great challenge mainly due to the special property of their natural habitat. Here, we design a new chemical proteomic probe OPA-S-S-alkyne that can efficiently and selectively target the lysines exposed on the cell surface and develop a chemical proteomics strategy for global analysis of surface functionality (GASF) in living cells. In total, we quantified 2639 cell surface lysines in Hela cell and several hundred residues with high reactivity were discovered, which represents the largest dataset of surface functional lysine sites to date. We discovered and validated that hyper-reactive lysine residues K382 on tyrosine kinase-like orphan receptor 2 (ROR2) and K285 on Endoglin (ENG/CD105) are at the protein interaction interface in co-crystal structures of protein complexes, emphasizing the broad potential functional consequences of cell surface lysines and GASF strategy is highly desirable for discovering new active and ligandable sites that can be functionally interrogated for drug discovery.
细胞表面蛋白在许多重要的生理过程中发挥着作用，也是药物靶点的主要类别，因为大多数治疗药物的作用对象是细胞表面的膜蛋白，以改变细胞信号转导。尽管意义重大，但针对膜蛋白的配体发现一直是一个巨大的挑战，主要是由于其天然栖息地的特殊性质。在这里，我们设计了一种新的化学蛋白质组学探针 OPA-S-S-alkyne，它可以有效地和选择性地针对细胞表面暴露的赖氨酸，开发了一种用于活细胞表面功能全局分析（GASF）的化学蛋白质组学策略。总的来说，我们在 Hela 细胞中定量了 2639 个细胞表面赖氨酸，发现了几百个具有高反应性的残基，这是迄今为止最大的表面功能赖氨酸位点数据集。我们发现并验证了酪氨酸激酶样孤儿受体 2（ROR2）上的高反应性赖氨酸残基 K382 和内皮糖蛋白（ENG/CD105）上的 K285 位于蛋白质相互作用界面，这在蛋白质复合物的共晶结构中得到了证实，强调了细胞表面赖氨酸的广泛潜在功能后果，GASF 策略对于发现新的活性和配体结合位点以进行药物发现是非常可取的。
