Native mass spectrometry (nMS) is becoming a crucial tool for analyzing membrane proteins (MPs), yet challenges remain in solubilizing and stabilizing their native conformations while resolving and characterizing the heterogeneity introduced by post-translational modifications and ligand binding. This review highlights recent advancements and persistent challenges in preparing MPs for nMS. Optimizing detergents and additives can significantly reduce sample heterogeneity and surface charge, enhancing MP signal quality and structural preservation in nMS. A strategic workflow incorporating affinity capture, stabilization agents, and size-exclusion chromatography to remove unfolded species demonstrates success in improving nMS characterization. Continued development of customized detergents and reagents tailored for specific MPs may further minimize heterogeneity and boost signals. Instrumental advances are also needed to elucidate more dynamically complex and labile MPs. Effective sample preparation workflows may provide insights into MP structures, dynamics, and interactions underpinning membrane biology. With ongoing methodological innovation, nMS shows promise to complement biophysical studies and facilitate drug discovery targeting this clinically important yet technically demanding protein class.

中文翻译：

---

制备用于非变性质谱的膜蛋白的进展和挑战


天然质谱 （nMS） 正在成为分析膜蛋白 （MP） 的重要工具，但在溶解和稳定其天然构象的同时，仍存在一些挑战，同时要解决和表征翻译后修饰和配体结合引入的异质性。本综述重点介绍了 MP 为 nMS 做准备的最新进展和持续挑战。优化去污剂和添加剂可以显著降低样品异质性和表面电荷，提高 MP 信号质量和 nMS 中的结构保留。结合亲和捕获、稳定剂和尺寸排阻色谱以去除未折叠物质的战略工作流程表明，该工作流程在改善 nMS 表征方面取得了成功。持续开发针对特定 MP 量身定制的去污剂和试剂可以进一步减少异质性并增强信号。还需要仪器进步来阐明动态更复杂和不稳定的 MP。有效的样品制备工作流程可以深入了解支撑膜生物学的 MP 结构、动力学和相互作用。通过不断的方法学创新，nMS 有望补充生物物理学研究，并促进针对这一临床重要但技术要求高的蛋白质类别的药物发现。