A delayed decoupling methyl-TROSY pulse sequence for atomic resolution studies of folded proteins and RNAs in condensates

IF 2 3区 化学 Q3 BIOCHEMICAL RESEARCH METHODS
Rashik Ahmed , Atul K. Rangadurai , Lisa Ruetz , Martin Tollinger , Christoph Kreutz , Lewis E. Kay
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引用次数: 0

Abstract

Solution NMR spectroscopy has tremendous potential for providing atomic resolution insights into the interactions between proteins and nucleic acids partitioned into condensed phases of phase-separated systems. However, the highly viscous nature of the condensed phase challenges applications, and in particular, the extraction of quantitative, site-specific information. Here, we present a delayed decoupling-based HMQC pulse sequence for methyl-TROSY studies of ‘client’ proteins and nucleic acids partitioned into ‘scaffold’ proteinaceous phase-separated solvents. High sensitivity and excellent quality spectra are recorded of a nascent form of superoxide dismutase and of a small RNA fragment partitioned into CAPRIN1 condensates.

Abstract Image

用于凝结物中折叠蛋白质和 RNA 原子分辨率研究的延迟去耦甲基-TROSY 脉冲序列
溶液核磁共振光谱具有巨大的潜力,可提供原子分辨率,深入了解分隔在相分离系统凝聚相中的蛋白质和核酸之间的相互作用。然而,凝聚相的高粘性给应用带来了挑战,尤其是在提取定量的特定位点信息方面。在此,我们介绍了一种基于延迟解耦的 HMQC 脉冲序列,用于对 "支架 "蛋白质相分离溶剂中的 "客户 "蛋白质和核酸进行甲基-TROSY 研究。研究人员记录了超氧化物歧化酶的新生形式和分入 CAPRIN1 凝聚物的小 RNA 片段的高灵敏度和高质量光谱。
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来源期刊
CiteScore
3.80
自引率
13.60%
发文量
150
审稿时长
69 days
期刊介绍: The Journal of Magnetic Resonance presents original technical and scientific papers in all aspects of magnetic resonance, including nuclear magnetic resonance spectroscopy (NMR) of solids and liquids, electron spin/paramagnetic resonance (EPR), in vivo magnetic resonance imaging (MRI) and spectroscopy (MRS), nuclear quadrupole resonance (NQR) and magnetic resonance phenomena at nearly zero fields or in combination with optics. The Journal''s main aims include deepening the physical principles underlying all these spectroscopies, publishing significant theoretical and experimental results leading to spectral and spatial progress in these areas, and opening new MR-based applications in chemistry, biology and medicine. The Journal also seeks descriptions of novel apparatuses, new experimental protocols, and new procedures of data analysis and interpretation - including computational and quantum-mechanical methods - capable of advancing MR spectroscopy and imaging.
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