Biological membranes are complex barriers in which membrane proteins and thousands of lipidic species participate in structural and functional interactions. Developing a strategic approach that allows uniform labeling of membrane proteins while maintaining a lipidic environment that retains functional interactions is highly desirable for in vitro fluorescence studies. Herein, we focus on complementing current methods by integrating the powerful processes of unnatural amino acid mutagenesis, bioorthogonal labeling, and the detergent-free membrane protein solubilization based on the amphiphilic styrene-maleic acid (SMA) polymer. Importantly, the SMA polymer preserves a thermodynamically stable shell of phospholipids. The approach that we present is both rapid and generalizable providing a population of uniquely labeled membrane proteins in lipid nanoparticles for quantitative fluorescence-based studies.

生物膜是复杂的屏障，其中膜蛋白和数千种脂质物质参与结构和功能相互作用。在体外，开发一种战略方法以允许对膜蛋白进行均匀标记，同时保持能够保留功能相互作用的脂质环境，这是非常需要的荧光研究。在这里，我们专注于通过结合基于两亲性苯乙烯-马来酸（SMA）聚合物的非天然氨基酸诱变，生物正交标记和无洗涤剂的膜蛋白增溶的强大过程来补充当前的方法。重要的是，SMA聚合物保留了磷脂的热力学稳定壳。我们提出的方法既快速又可推广，可在脂质纳米颗粒中提供一组独特标记的膜蛋白，用于基于荧光的定量研究。