超极化的核磁共振用于蛋白质 - 连接体相互作用的表征
Youngbok Lee1, Haifeng Zeng, Simon Ruedisser
1Center for Biological NMR, Department of Chemistry, Texas A&M University, College Station, Texas 77843, USA.
Journal of the American Chemical Society
|October 2, 2012
概括
动态核极化增强了的NMR灵敏度,使得低度蛋白质-连接体相互作用的检测成为药物发现的关键. 这种技术可以改善对候选药物的选,即使是那些具有较弱结合亲和力的药物.
科学领域:
- 生物化学 生物化学
- 化学生物学 化学生物学
- 药物发现 药物发现 药物发现
背景情况:
- NMR光谱对于研究药物发现中的蛋白质-连接体相互作用是有价值的,这是由于许多药物中的简单光谱和的存在.
- 核磁共振的一个主要限制是它的低灵敏度,需要高的蛋白质度和配体:蛋白质比率.
- 对化小分子的敏感检测对于有效的药物查至关重要.
研究的目的:
- 提出利用动态核极化 (DNP) 的技术,以增强 (19) F NMR信号,用于检测蛋白质 - 配体相互作用.
- 证明DNP增强型NMR的应用,用于分析强,中,弱结合模式的配体.
- 探索DNP增强型NMR作为替代药物发现查方法的潜力.
主要方法:
- 利用动态核极化,在19F NMR中实现数千倍的信号增强.
- 应用增强的NMR技术来检测化小分子的亚微分子度.
- 开发方法来确定解离常数,并分析不同结合强度和交换模式中的配体.
主要成果:
- 取得了显著的 (19) F信号增强,使得能够检测出亚微粒度联体度.
- 成功地应用了该技术在强,中等和弱结合场景中研究蛋白质 - 连接体相互作用.
- 证明了在缓慢交换中检测配体的能力,这对传统的NMR方法来说具有挑战性.
结论:
- 动态核极化显著提高 (19) 蛋白质-配体相互作用研究的NMR灵敏度.
- 这种增强的灵敏度可以检测低度的配体,并描述以前难以评估的结合相互作用.
- 用DNP增强的NMR提供了更快,更有信息的方法,为药物发现查提供了一个有希望的替代方案.
相关概念视频
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...
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...
Atomic Nuclei: Magnetic Resonance
The number of nuclear spins aligned in the lower energy state is slightly greater than those in the higher energy state. In the presence of an external magnetic field, as the spins precess at the Larmor frequency, the excess population results in a net magnetization oriented along the z axis. When a pulse or a short burst of radio waves at the Larmor frequency is applied along the x axis, the coupling of frequencies causes resonance and flips the nuclear spins of the excess population from the...
Protein Dynamics in Living Cells
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Nuclear Overhauser Enhancement (NOE)
Irradiation of a spin-active nucleus causes an increase or decrease in the signal intensity of neighboring nuclei that are not necessarily chemically bonded or involved in J-coupling. This phenomenon, called the nuclear Overhauser enhancement (NOE), results from through-space interactions between the nuclear spins. The NOE effect decreases with increasing internuclear distance and is generally not observed beyond 4 angstroms. In NOE, dipole-dipole interactions between neighboring spin-active...


