The SOFAST-HMBC-HMQC experiment for pairing geminal methyl groups in valine and leucine side-chains

IF 1.9 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Journal of Biomolecular NMR Pub Date : 2025-04-07 DOI:10.1007/s10858-025-00464-z
Ana Paula Aguilar Alva, Lucas Siemons, Ulric B. le Paige, Coline Wiame, Florence Cordier, Nicolas Wolff, Guillaume Bouvignies, Philippe Pelupessy, Fabien Ferrage
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Abstract

Methyl groups are essential probes for characterising interactions and dynamics in large proteins. HN-based triple-resonance NMR experiments are often too insensitive for methyl assignments, making a NOESY-based approach an efficient strategy. Linking geminal methyl groups in leucine and valine residues is a crucial step in such NOESY-based methyl resonance assignment strategies. This link can be established unambiguously with the 3D-HMBC-HMQC experiment, introduced for large U-[12C, 2H] LV-[13CH3]2-labelled proteins. Here, we introduce the SOFAST variant of the 3D-HMBC-HMQC experiment which provides spectra with fewer artefacts arising from the water signal and a mean increase in signal-to-noise ratio per unit time of 16% compared to the original experiment with an optimised recovery delay.

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缬氨酸和亮氨酸侧链上双甲基配对的SOFAST-HMBC-HMQC实验。
甲基是表征大蛋白质相互作用和动力学的必要探针。基于hn的三共振核磁共振实验通常对甲基分配过于不敏感,使得基于nosy的方法成为一种有效的策略。连接亮氨酸和缬氨酸残基的双甲基是这种基于noes的甲基共振分配策略的关键步骤。这种联系可以通过3D-HMBC-HMQC实验明确地建立起来,该实验引入了大型U-[12C, 2H] LV-[13CH3]2标记的蛋白质。在这里,我们介绍了3D-HMBC-HMQC实验的SOFAST变体,该实验提供的光谱中由水信号产生的伪影更少,与优化的恢复延迟的原始实验相比,单位时间的信噪比平均增加了16%。
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来源期刊
Journal of Biomolecular NMR
Journal of Biomolecular NMR 生物-光谱学
CiteScore
6.00
自引率
3.70%
发文量
19
审稿时长
6-12 weeks
期刊介绍: The Journal of Biomolecular NMR provides a forum for publishing research on technical developments and innovative applications of nuclear magnetic resonance spectroscopy for the study of structure and dynamic properties of biopolymers in solution, liquid crystals, solids and mixed environments, e.g., attached to membranes. This may include: Three-dimensional structure determination of biological macromolecules (polypeptides/proteins, DNA, RNA, oligosaccharides) by NMR. New NMR techniques for studies of biological macromolecules. Novel approaches to computer-aided automated analysis of multidimensional NMR spectra. Computational methods for the structural interpretation of NMR data, including structure refinement. Comparisons of structures determined by NMR with those obtained by other methods, e.g. by diffraction techniques with protein single crystals. New techniques of sample preparation for NMR experiments (biosynthetic and chemical methods for isotope labeling, preparation of nutrients for biosynthetic isotope labeling, etc.). An NMR characterization of the products must be included.
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