Strategies for Protein NMR in Escherichia coli

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	Strategies for Protein NMR in Escherichia coli
	Xu, Guohua 1; Ye, Yansheng 1,2; Liu, Xiaoli 1; Cao, Shufen 1; Wu, Qiong 1; Cheng, Kai 1,2; Liu, Maili 1; Pielak, Gary J.3,4; Li, Conggang 1
刊名	BIOCHEMISTRY
	2014-04-01
卷号	53 期号:12 页码:1971-1981
英文摘要	In-cell NMR spectroscopy provides insight into protein conformation, dynamics, and function at atomic resolution in living cells. Systematic evaluation of isotopic-labeling strategies is necessary to observe the target protein in the sea of other molecules in the cell. Here, we investigate the detectability, sensitivity, and resolution of in-cell NMR spectra of the globular proteins GB1, ubiquitin, calmodulin, and bcl-xl-cutloop, resulting from uniform N-15 enrichment (with and without deuteration), selective N-15-Leu enrichment, C-13-methyl enrichment of isoleucine, leucine, valine, and alanine, fractional C-13 enrichment, and F-19 labeling. Most of the target proteins can be observed by F-19 labeling and C-13 enrichment with direct detection because selectively labeling suppresses background signals and because deuteration improves in-cell spectra. Our results demonstrate that the detectability of proteins is determined by weak interactions with intercellular components and that choosing appropriate labeling strategies is critical for the success of in-cell protein NMR studies.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biochemistry & Molecular Biology
研究领域[WOS]	Biochemistry & Molecular Biology
关键词[WOS]	IN-CELL NMR ; AROMATIC SIDE-CHAINS ; LIVING CELLS ; F-19 NMR ; MULTIDIMENSIONAL NMR ; METHYL-GROUPS ; STINT-NMR ; SPECTROSCOPY ; STABILITY ; DYNAMICS
收录类别	SCI
语种	英语
WOS记录号	WOS:000333776300010
公开日期	2015-07-14
内容类型	期刊论文
源URL	[http://ir.wipm.ac.cn/handle/112942/1366]
专题	武汉物理与数学研究所_磁共振基础研究部
作者单位	1.Chinese Acad Sci, Wuhan Inst Phys & Math, Ctr Magnet Resonance,Key Lab Magnet Resonance Bio, State Key Lab Magnet Resonance & Atom & Mol Phys, Wuhan 430071, Peoples R China 2.Chinese Acad Sci, Grad Univ, Beijing 100029, Peoples R China 3.Univ N Carolina, Dept Chem, Chapel Hill, NC 27599 USA 4.Univ N Carolina, Dept Biochem & Biophys, Chapel Hill, NC 27599 USA
推荐引用方式 GB/T 7714	Xu, Guohua,Ye, Yansheng,Liu, Xiaoli,et al. Strategies for Protein NMR in Escherichia coli[J]. BIOCHEMISTRY,2014,53(12):1971-1981.
APA	Xu, Guohua.,Ye, Yansheng.,Liu, Xiaoli.,Cao, Shufen.,Wu, Qiong.,...&Li, Conggang.(2014).Strategies for Protein NMR in Escherichia coli.BIOCHEMISTRY,53(12),1971-1981.
MLA	Xu, Guohua,et al."Strategies for Protein NMR in Escherichia coli".BIOCHEMISTRY 53.12(2014):1971-1981.