Lysine methylation: A strategy to improve in-cell NMR spectroscopy of proteins

被引:0
作者
Xiao, Xiong [1 ,2 ]
Zhan, Jianhua [1 ,2 ]
Liu, Biao [1 ,3 ]
Zhu, Qinjun [1 ]
Wang, Guan [1 ]
Zeng, Danyun [1 ,2 ]
Liu, Caixiang [1 ,2 ]
Jiang, Bin [1 ,2 ,3 ]
He, Lichun [1 ,2 ]
Gong, Zhou [1 ,2 ]
Zhou, Xin [1 ,2 ,3 ,4 ]
Zhang, Xu [1 ,2 ,3 ,4 ]
Liu, Maili [1 ,2 ,3 ,4 ]
机构
[1] Chinese Acad Sci, Key Lab Magnet Resonance Biol Syst, State Key Lab Magnet Resonance & Atom & Mol Phys, Natl Ctr Magnet Resonance Wuhan,Wuhan Inst Phys &, Wuhan 430071, Peoples R China
[2] Univ Chinese Acad Sci, Beijing 100049, Peoples R China
[3] Huazhong Univ Sci & Technol, Wuhan Natl Lab Optoelect, Wuhan 430074, Peoples R China
[4] Opt Valley Lab, Wuhan 430074, Peoples R China
基金
中国国家自然科学基金;
关键词
In-cell NMR; Intracellular environment; Non-specific interactions; Lysine methylation Signal enhancement; CYTOCHROME-C; REDUCTIVE METHYLATION; MULTIDIMENSIONAL NMR; F-19; NMR; EXPRESSION; DOMAIN; TOOL;
D O I
10.1016/j.aca.2024.343099
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Background: In-cell NMR is a valuable technique for investigating protein structure and function in cellular environments. However, challenges arise due to highly crowded cellular environment, where nonspecific interactions between the target protein and other cellular components can lead to signals broadening or disappearance in NMR spectra. Results: We implemented chemical reduction methylation to selectively modify lysine residues on protein surfaces aiming to weaken charge interactions and recover obscured NMR signals. This method was tested on six proteins varying in molecular size and lysine content. While methylation did not disrupt the protein's native conformation, it successful restored some previously obscured in-cell NMR signals, particularly for proteins with high isoelectric points that decreased post-methylation. Significance: This study affirms lysine methylation as a feasible approach to enhance the sensitivity of in-cell NMR spectra for protein studies. By mitigating signal loss due to nonspecific interactions, this method expands the utility of in-cell NMR for investigating proteins in their natural cellular environment, potentially leading to more accurate structural and functional insights.
引用
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页数:9
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