用19F NMR鉴别GPCR的构象动力学和结合动力学

IF 4 3区 医学 Q1 PHARMACOLOGY & PHARMACY
R.S. Prosser , Nicholas A. Alonzi
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引用次数: 0

摘要

19F NMR提供了一种从集合的角度监测G蛋白偶联受体(GPCR)构象动力学的方法。虽然X射线晶体学提供了特定状态的精细解析的高分辨率结构,但它通常不能概括功能状态的真正集合。氟(19F)NMR为构象系综提供了一个高度灵敏的光谱窗口,通常允许直接定量可分辨状态。此外,基于T1和T2的直接弛豫实验允许在纳秒到秒的时间尺度上研究给定状态内的波动和状态之间的交换。方便的是,大多数生物系统都不含氟。因此,通过氟化氨基酸类似物或硫醇反应性氟化标签,F或CF3报告子可以位点特异性地结合到感兴趣的蛋白质中。在这篇综述中,将从小分子NMR(即药物或小分子与受体的相互作用)或大分子NMR(如受体和受体-G蛋白复合物的构象动力学)的角度介绍氟标记方案和19F NMR实验。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Discerning conformational dynamics and binding kinetics of GPCRs by 19F NMR

19F NMR provides a way of monitoring conformational dynamics of G-protein coupled receptors (GPCRs) from the perspective of an ensemble. While X-ray crystallography provides exquisitely resolved high-resolution structures of specific states, it generally does not recapitulate the true ensemble of functional states. Fluorine (19F) NMR provides a highly sensitive spectroscopic window into the conformational ensemble, generally permitting the direct quantification of resolvable states. Moreover, straightforward T1- and T2-based relaxation experiments allow for the study of fluctuations within a given state and exchange between states, on timescales spanning nanoseconds to seconds. Conveniently, most biological systems are free of fluorine. Thus, via fluorinated amino acid analogues or thiol-reactive fluorinated tags, F or CF3 reporters can be site specifically incorporated into proteins of interest. In this review, fluorine labeling protocols and 19F NMR experiments will be presented, from the perspective of small molecule NMR (i.e. drug or small molecule interactions with receptors) or macromolecular NMR (i.e. conformational dynamics of receptors and receptor–G-protein complexes).

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来源期刊
CiteScore
8.80
自引率
2.50%
发文量
131
审稿时长
4-8 weeks
期刊介绍: Current Opinion in Pharmacology (COPHAR) publishes authoritative, comprehensive, and systematic reviews. COPHAR helps specialists keep up to date with a clear and readable synthesis on current advances in pharmacology and drug discovery. Expert authors annotate the most interesting papers from the expanding volume of information published today, saving valuable time and giving the reader insight on areas of importance.
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