The molecular nanoscale organization of the surfaceome is a fundamental regulator of cellular signaling in health and disease. Technologies for mapping the spatial relationships of cell surface receptors and their extracellular signaling synapses would unlock theranostic opportunities to target protein communities and the possibility to engineer extracellular signaling. Here, we develop an optoproteomic technology termed LUX-MS that enables the targeted elucidation of acute protein interactions on and in between living cells using light-controlled singlet oxygen generators (SOG). By using SOG-coupled antibodies, small molecule drugs, biologics and intact viral particles, we demonstrate the ability of LUX-MS to decode ligand receptor interactions across organisms and to discover surfaceome receptor nanoscale organization with direct implications for drug action. Furthermore, by coupling SOG to antigens we achieved light-controlled molecular mapping of intercellular signaling within functional immune synapses between antigen-presenting cells and CD8⁺ T cells providing insights into T cell activation with spatiotemporal specificity. LUX-MS based decoding of surfaceome signaling architectures thereby provides a molecular framework for the rational development of theranostic strategies.

表面组的分子纳米级组织是健康和疾病中细胞信号传导的基本调节剂。绘制细胞表面受体及其细胞外信号突触的空间关系的技术将解锁靶向蛋白质群落的治疗诊断机会和设计细胞外信号传导的可能性。在这里，我们开发了一种称为 LUX-MS 的光蛋白质组学技术，该技术能够使用光控单线态氧发生器 (SOG) 有针对性地阐明活细胞上和活细胞之间的急性蛋白质相互作用。通过使用 SOG 偶联抗体、小分子药物、生物制剂和完整的病毒颗粒，我们证明了 LUX-MS 能够解码生物体之间的配体受体相互作用，并发现对药物作用有直接影响的表面组受体纳米级组织。此外，通过将 SOG 与抗原偶联，我们在抗原呈递细胞和 CD8 之间的功能性免疫突触内实现了细胞间信号的光控分子定位。⁺  T 细胞提供对具有时空特异性的 T 细胞活化的见解。因此，基于 LUX-MS 的表面组信号结构解码为治疗诊断策略的合理发展提供了分子框架。