相关实验视频

Updated: Jun 17, 2026

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

High Precision FRET at Single-molecule Level for Biomolecule Structure Determination

Published on: May 13, 2017

在大肠杆菌中的蛋白质 (19) F NMR

Conggang Li¹, Gui-Fang Wang, Yaqiang Wang

¹Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.

Journal of the American Chemical Society

|January 7, 2010

概括

核磁共振 (NMR) 光谱现在可以检测活细菌细胞内较大的蛋白质. 这一进步使用-19 (19F) 标签,克服了传统-15 (15N) 丰富的局限性,用于观察球状蛋白质.

科学领域:

生物化学生物化学
生物物理学的生物物理.
结构生物学结构生物学

背景情况:

-15 (15N) 丰富有助于观察大肠杆菌中的无序蛋白质,但对于大多数球状蛋白质来说不足.
在检测使用细胞内NMR的无序与球状蛋白质方面存在分歧,限制了更广泛的应用.
目前的方法很难在细胞环境中检测到更大的球状蛋白质.

研究的目的:

克服现有的细胞内NMR技术检测球状蛋白质的局限性.
扩大细胞内NMR研究蛋白质结构和动态的能力.
为了能够在活细胞中观察更大的蛋白质,无论是无序的还是球状的.

主要方法:

使用-19 (19F) 标记蛋白质与特定的氨基酸类似物.
用于标记较小的球状蛋白质的3 - - 铁氨酸 (约. 10kDa) 的时间.
加入三甲基-L-氨,用于标记较大的球状蛋白质 (高达100kDa).

主要成果:

在细胞中成功观察到标有3 - - 铁素的小球状蛋白质的共振.
通过使用trifluoromethyl-L-phenylalanine克服了对较大的蛋白质的共振扩展问题.
证明了获得细胞内部球状和无序蛋白质的特定位置结构和动态信息的能力.

结论:

用适当的标记策略进行-19 (19F) 核磁共振显著提高了活细胞内球状蛋白质的检测.
这种方法克服了以前的尺寸限制,使细胞内核磁共振能用于研究蛋白质行为的更广泛的应用.
对蛋白质的特定位置结构和动态洞察力现在可以在体内实现,无论是无序的还是球状蛋白质类型.

更多相关视频

15N CPMG Relaxation Dispersion for the Investigation of Protein Conformational Dynamics on the µs-ms Timescale

¹⁵N CPMG Relaxation Dispersion for the Investigation of Protein Conformational Dynamics on the µs-ms Timescale

Published on: April 19, 2021

The Multifaceted Benefits of Protein Co-expression in Escherichia coli

The Multifaceted Benefits of Protein Co-expression in Escherichia coli

Published on: February 5, 2015

相关概念视频

Other Nuclides: 31P, 19F, 15N NMR

Other Nuclides: ³¹P, ¹⁹F, ¹⁵N NMR

Many organic, inorganic, and biological molecules contain spin-half nuclei such as nitrogen-15, fluorine-19, and phosphorus-31. As a result, NMR studies of these nuclei have found extensive applications in chemical and biological research.
While fluorine-19 and phosphorous-31 have high natural abundances (100%) and positive gyromagnetic ratios, nitrogen-15 has a low natural abundance and a negative gyromagnetic ratio. However, nitrogen-15 is still preferred over nitrogen-14 (which has a high...

Stringent Response in E. coli

Stringent Response in E. coli

Bacterial growth is closely tied to nutrient availability, with cells proliferating exponentially under favorable conditions and entering a stationary phase when resources become scarce. This transition is mediated by a regulatory mechanism known as the stringent response, which allows bacteria to adapt to nutrient deprivation by modulating gene expression and metabolic activity.During nutrient scarcity, intracellular amino acid levels decline. It results in the accumulation of uncharged tRNAs...

Single-Strand DNA Binding Proteins

Single-Strand DNA Binding Proteins

For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...