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相关概念视频

Nuclear Magnetic Resonance (NMR): Overview01:07

Nuclear Magnetic Resonance (NMR): Overview

8.1K
Nuclear magnetic resonance (NMR) is a phenomenon exhibited by certain nuclei that can absorb characteristic radio frequency radiation under certain conditions. NMR has been extensively applied in molecular spectroscopy and medical diagnostic imaging. In both these applications, the molecule or subject under study is placed in a magnetic field and irradiated with radio frequency energy.
NMR spectroscopy generates a spectrum where the characteristic absorption frequencies of the sample are...
8.1K
NMR Spectrometers: Overview01:20

NMR Spectrometers: Overview

2.5K
NMR spectrometers consist of a strong magnet, a radiofrequency transmitter, and a detector attached to a computer console for recording spectra of samples containing NMR-active nuclei. In first-generation NMR instruments called continuous-wave spectrometers, the resonance frequencies of the nuclei are determined by frequency-sweep or field-sweep methods. The magnetic field strength is fixed and the rf signal is swept in the former, while the radiofrequency signal is fixed and the magnetic field...
2.5K
¹H NMR: Interpreting Distorted and Overlapping Signals01:02

¹H NMR: Interpreting Distorted and Overlapping Signals

1.7K
Spin systems where the difference in chemical shifts of the coupled nuclei is greater than ten times J are called first-order spin systems. These nuclei are weakly coupled, and their chemical shifts and coupling constant can generally be estimated from the well-separated signals in the spectrum.
As Δν decreases and the signals move closer, the doublets appear increasingly distorted. The intensities of the inner lines increase at the cost of those of the outer lines as the signals are...
1.7K
Two-Dimensional (2D) NMR: Overview01:12

Two-Dimensional (2D) NMR: Overview

1.8K
The 1D NMR spectrum of large and complex molecules like natural products has complicated splitting patterns and overlapping signals, which can be easily interpreted using 2-dimensional (2D) NMR. Unlike 1D NMR, 2D NMR has two frequency axes that provide the coupling information between the nucleus A and nucleus B in a molecule. The process from which 2D spectra are obtained has four steps.
The first step is the preparation period, during which nucleus A is excited with a radiofrequency pulse....
1.8K
NMR Spectrometers: Radiofrequency Pulses and Pulse Sequences01:17

NMR Spectrometers: Radiofrequency Pulses and Pulse Sequences

2.1K
A pulse is a short burst of radio waves distributed over a range of frequencies that simultaneously excites all the nuclei in the sample. Upon passing a radio frequency pulse along the x-axis, the nuclei absorb energy corresponding to their Larmor frequencies and achieve resonance. This shifts the net magnetization vector from the z-axis toward the transverse plane. This angle of rotation of the magnetization vector, or the flip angle, is proportional to the duration and intensity of the pulse.
2.1K
Applications Of NMR In Biology01:25

Applications Of NMR In Biology

4.7K
Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics  for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.
4.7K

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Directed Evolution Method in Saccharomyces cerevisiae: Mutant Library Creation and Screening
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通过NMR指导的定向进化

Sagar Bhattacharya1, Eleonora G Margheritis2, Katsuya Takahashi2

  • 1Department of Chemistry, Syracuse University, Syracuse, NY, USA.

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|October 5, 2022
PubMed
概括
此摘要是机器生成的。

研究人员开发了一种新的NMR光谱法来识别蛋白质中的关键突变部位. 这种方法有效地将肌球蛋白转化为Kemp消除酶,仅用三种突变,展示了蛋白质工程的强大工具.

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科学领域:

  • 生物化学和分子生物学
  • 蛋白质工程
  • 酶催化

背景情况:

  • 定向进化对于蛋白质的改进和功能化至关重要,但受到巨大的序列空间的限制.
  • 目前用于预测有益突变的方法通常依赖于结构或生物信息学数据,这些数据并不总是可用.
  • 鉴定远离活性部位的突变,这些突变可以显著增强酶特性,仍然具有挑战性.

研究的目的:

  • 建立一种使用NMR光谱的新方法来识别酶中的突变热点.
  • 证明这种方法在设计新型酶功能中的实用性.
  • 克服蛋白质工程现有预测方法的局限性.

主要方法:

  • 使用核磁共振 (NMR) 光谱识别突变热点.
  • 应用了一项涉及肌球蛋白定向演变的概念验证研究.
  • 引入了最小的突变来赋予新的酶活性.

主要成果:

  • 通过仅使用三种突变,成功将非酶蛋白转化为高效的Kemp消除酶.
  • 达到与自然存在的酶相匹配的化效率水平.
  • 证明这种方法在效率方面优于当前的蛋白质设计方法.

结论:

  • 核磁共振光谱提供了一种简单有效的实验方法来识别蛋白质工程中的关键残留物.
  • 这种方法绕过了先验结构或生物信息信息的需求,提高了适用性.
  • 这种方法具有显著的潜力,