通过膜内化学修饰（IMCM）探测 G 蛋白偶联受体跨膜域的溶剂可及性。
Solvent accessibility of a GPCR transmembrane domain probed by in-membrane chemical modification (IMCM).
机构信息
iHuman Institute, ShanghaiTech University, China.
School of Life Science and Technology, ShanghaiTech University, China.
出版信息
FEBS Lett. 2023 Jun;597(11):1541-1549. doi: 10.1002/1873-3468.14627. Epub 2023 Apr 29.
G protein-coupled receptors (GPCRs) transmit signals from drugs across cell membranes, leading to associated physiological effects. To study the structural basis of the transmembrane signalling, in-membrane chemical modification (IMCM) has previously been introduced for F-labelling of GPCRs expressed in Spodoptera frugiperda (Sf9) insect cells. Here, IMCM is used with the A adenosine receptor (A AR) expressed in Pichia pastoris; F-NMR revealed nearly complete solvent protection of the A AR transmembrane domain in the membrane and in 2,2-didecylpropane-1,3-bis-β-D-maltopyranoside (LMNG)/cholesteryl hemisuccinate (CHS) micelles, and extensive solvent accessibility for A AR in n-dodecyl β-D-maltoside (DDM)/CHS micelles. No Cys residue dominated non-specific labelling with 2,2,2-trifluoroethanethiol. These observations yield an improved protocol for IMCM F-labelling of GPCRs and new insights into variable solvent accessibility for function-related characterization of GPCRs.
G 蛋白偶联受体（GPCRs）将药物信号从细胞膜传递到细胞内，引发相关的生理效应。为了研究跨膜信号传递的结构基础，先前已经引入了细胞膜内化学修饰（IMCM），用于标记在 Spodoptera frugiperda（ Sf9）昆虫细胞中表达的 GPCRs。在这里，将 IMCM 用于在毕赤酵母中表达的 A 腺苷受体（A AR）； 19 F-NMR 显示 A AR 跨膜结构域在膜中和 2,2-二癸基丙-1,3-双-β-D-麦芽糖苷（LMNG）/胆固醇琥珀酸酯（CHS）胶束中几乎完全被溶剂保护，而在正十二烷基 β-D-麦芽糖苷（DDM）/CHS 胶束中 A AR 具有广泛的溶剂可及性。没有 Cys 残基主导非特异性用 2,2,2-三氟乙硫醇标记。这些观察结果为 GPCRs 的 IMCM 19 F 标记提供了改进的方案，并为 GPCRs 的功能相关表征的可变溶剂可及性提供了新的见解。
Zotero 插件