单分子双色探针用于脂微区动态可视化及脂微区与K-Ras蛋白相互作用机理研究

脂微区有序相和无序相的形成与重排启动了一系列重要的生理及病理学进程，如细胞的增殖分化、内吞、免疫应答和病原体入侵。细胞可通过对脂微区的重排，实现对细胞膜张力的调控及特定蛋白的富集与解离，从而完成细胞形变、信号传导与跨膜转运过程的调控。而揭示脂微区的动态过程和翻译后修饰蛋白与脂微区相互作用的规律，有助于推动对细胞定向迁移、癌细胞扩散抑制的深入研究。N’N-二芳基-二氢二苯并吩嗪是一种基于“振动诱导发光（Vibration-Induced-Emission, VIE）”机制的新型单分子比率式双色荧光染料，对所处环境的极性和粘度改变高度敏感，是脂微区动态过程与蛋白相互作用研究的理想探针。本项目拟发展基于VIE机制的双色荧光探针，旨在实现脂微区的动态可视化，揭示K-Ras蛋白与脂微区相互作用的选择性机制及内在规律，为细胞膜应力改变、细胞定向迁移、癌细胞扩散抑制的深入理解提供新的研究策略。

The complex nature of cell plasma membranes raised intensive research in the last decades, stimulated by the hypothesis of lipid microdomains. Cellular membranes are laterally heterogeneous and consist of transient and dynamic domains with varying properties, which prominently include liquid-disordered and liquid-ordered domains. Membrane domains can be induced and regulated by a variety of interactions to initiates a range of central biological and pathological events, which include specific lipid–lipid and lipid–protein interactions, and interactions between membrane components and the underlying cytoskeleton. The rearrangement of lipid microdomains can regulate cellular deformation, signal transduction and transmembrane transport processes through modulation of membrane tension,association and dissociation of membrane proteins. N,N’‑disubstituted- dihydrodibenzo[a,c]phenazines is a novel ratiometric dual-emission fluorescence probes based on vibration-induced-emission (VIE) mechanism, which is highly sensitive to changes in polarity and viscosity, and is an ideal probe for studying the dynamic processes of association between membrane proteins with lipid microdomains. This project employs a series of dual-emission unimolecular probe to visualize the dynamics change of lipid microdomains, aims to unravel the basis of intrinsic interaction between K-Ras protein and lipid microdomains, and provides fundamental insights into the physiology of cell membrane stress change, cell directional-migration, and inhibition of cancer cell spreading.