Marassi Francesca M, Pintacuda Guido
Department of Biophysics, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226-3548, USA.
Centre de RMN à Très Hauts Champs de Lyon, CNRS / ENS Lyon / UCB Lyon 1, 5 rue de la Doua, 69100 Villeurbanne, France.
Curr Opin Struct Biol. 2025 Aug 8;94:103129. doi: 10.1016/j.sbi.2025.103129.
Membrane proteins have evolved to function as part of specialized biological membranes, and their structures and activities are highly susceptible to their local environment. Detergents and lipid mimetics replicate certain aspects of biological membranes, and have been used to produce an exceptional body of structural data, but do not fully capture the complex, asymmetric properties of the native environment and can alter structure and function. Here, we review recent advances in nuclear magnetic resonance (NMR) that enable the examination of membrane protein structure and activity in situ, within native membranes. The development of optimized protein expression strategies, isotopic labeling schemes, powerful instrumentation and specialized pulse sequences offer new opportunities for exploring the new frontier of in situ structural biology. By outlining the framework for in situ NMR of membrane proteins from conceptualization to experiments we hope to inspire new research in this growing and important area.
膜蛋白已进化为专门生物膜的一部分发挥功能，其结构和活性对局部环境高度敏感。去污剂和脂质模拟物可复制生物膜的某些方面，并已用于产生大量出色的结构数据，但无法完全捕捉天然环境的复杂、不对称特性，且可能改变结构和功能。在此，我们综述了核磁共振（NMR）的最新进展，这些进展能够在天然膜内原位检测膜蛋白的结构和活性。优化的蛋白质表达策略、同位素标记方案、强大的仪器设备和专门的脉冲序列的发展为探索原位结构生物学的新领域提供了新机会。通过概述从概念化到实验的膜蛋白原位NMR框架，我们希望激发这一不断发展且重要领域的新研究。
