Department of Materials, Department of Bioengineering, and Institute of Biomedical Engineering, Imperial College London, London, SW7 2AZ, United Kingdom.
Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, 17177, Sweden.
Angew Chem Int Ed Engl. 2024 Apr 2;63(14):e202314786. doi: 10.1002/anie.202314786. Epub 2024 Mar 4.
Due to the variety of roles served by the cell membrane, its composition and structure are complex, making it difficult to study. Bioorthogonal reactions, such as the strain promoted azide-alkyne cycloaddition (SPAAC), are powerful tools for exploring the function of biomolecules in their native environment but have been largely unexplored within the context of lipid bilayers. Here, we developed a new approach to study the SPAAC reaction in liposomal membranes using azide- and strained alkyne-functionalized Förster resonance energy transfer (FRET) dye pairs. This study represents the first characterization of the SPAAC reaction between diffusing molecules inside liposomal membranes. Potential applications of this work include in situ bioorthogonal labeling of membrane proteins, improved understanding of membrane dynamics and fluidity, and the generation of new probes for biosensing assays.
由于细胞膜具有多种功能，其组成和结构较为复杂，因此难以对其进行研究。生物正交反应，如应变促进的叠氮化物-炔烃环加成（SPAAC）反应，是在天然环境中探索生物分子功能的有力工具，但在脂质双层中尚未得到广泛应用。在这里，我们开发了一种新方法，使用叠氮化物和应变炔基功能化的Förster 共振能量转移（FRET）染料对来研究脂质体膜中的 SPAAC 反应。本研究首次对脂质体膜内扩散分子之间的 SPAAC 反应进行了表征。这项工作的潜在应用包括对膜蛋白的原位生物正交标记、对膜动力学和流动性的更好理解，以及为生物传感测定生成新的探针。
