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相关概念视频

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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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...
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IR Spectrometers01:25

IR Spectrometers

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There are two main infrared (IR) spectrophotometers: dispersive IR spectrometers and Fourier transform infrared (FTIR) spectrometers. In a dispersive IR spectrometer, a beam of infrared radiation produced by a hot wire is divided into two parallel equal-intensity beams using mirrors. One beam passes through the sample, while another is a reference beam. The beams then move through the monochromator, which separates the radiations into a continuous spectrum of different frequencies. The...
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Infrared (IR) Spectroscopy: Overview01:09

Infrared (IR) Spectroscopy: Overview

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When electromagnetic radiation passes through a material, atoms or molecules transition from a lower to a higher energy state by absorbing radiation corresponding to the energy difference between the two states. The absorption of infrared (IR) radiation causes transitions between vibrational energy levels in a molecule. Therefore, IR spectroscopy is a useful analytical tool for determining the molecular structure of molecules.
Different compounds display unique properties due to their...
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IR Frequency Region: Fingerprint Region01:03

IR Frequency Region: Fingerprint Region

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IR spectra are divided into two main regions: the diagnostic region and the fingerprint region. The diagnostic region of the spectrum lies above 1500 cm−1. The absorptions resulting from single-bond vibrations of the N–H, C–H, and O–H stretch at higher wavenumbers and appear on the left side of the spectrum. The stretching absorptions of the C≡C and C≡N occur between 2100–2300 cm−1. In contrast, those arising from stretching absorptions of the...
907
IR Spectroscopy: Molecular Vibration Overview01:24

IR Spectroscopy: Molecular Vibration Overview

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When Infrared (IR) radiation passes through a covalently bonded molecule, the bonds transition from lower to higher vibrational levels. The fundamental vibrational motions that result in infrared absorption can be classified as stretching or bending vibrations.
Stretching vibrations are vibrational motions that occur along the bond line, changing the bond length or distance between two bonded atoms. They are further distinguished as symmetric or asymmetric. In symmetric stretching, the...
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Applications of IR Spectroscopy: Overview01:11

Applications of IR Spectroscopy: Overview

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The non-destructive nature and ability to provide valuable chemical information make IR spectroscopy a versatile technique with broad applications in various scientific and industrial fields. IR spectroscopy is commonly used to identify and characterize organic and inorganic compounds. It provides information about the functional groups present in a molecule and the bonding between atoms. This helps in the structural elucidation of compounds during organic synthesis, pharmaceutical research,...
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Updated: Jul 9, 2025

Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy
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使用FT-IR光谱法确定蛋白质动态的协议.

Hao Shen1, Cuiping Fu2, Junting Zhang2

  • 1Institute of Mass Spectrometry, School of Material Science and Chemical Engineering, Ningbo University, Ningbo, Zhejiang 315211, China; State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Ningbo University, Ningbo, 315211, China.

STAR protocols
|December 3, 2023
PubMed
概括
此摘要是机器生成的。

这项研究引入了一个里埃变换红外光谱 (FT-IR) 协议,用于测量使用胺/ (H/D) 交换的蛋白质动态. 这种方法有助于分析突变或相互作用如何影响蛋白质H/D的汇率.

关键词:
生物物理学的生物物理.蛋白质生物化学 蛋白质生物化学结构生物学 结构生物学

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科学领域:

  • 生物化学 生物化学
  • 频谱学是一种光谱学.
  • 结构生物学 结构生物学

背景情况:

  • 蛋白质功能与动态结构变化密切相关.
  • 蛋白质动态对于生物过程至关重要,可以使用胺/ (H/D) 交换来研究.
  • 胺H/D交换是一种强大的技术,用于探测蛋白质的灵活性和构造性景观.

研究的目的:

  • 提出评估蛋白质动态的详细方案.
  • 建立一种利用里埃变换红外光谱 (FT-IR) 来进行H/D交换分析的方法.
  • 为研究各种因素对蛋白质动态的影响提供一个框架.

主要方法:

  • 样品制备蛋白质. 样品制备蛋白质.
  • 福里埃变换红外 (FT-IR) 光谱收集.
  • 对FT-IR光谱进行详细的分析程序,以量化H/D交换.

主要成果:

  • 该协议可以通过FT-IR光谱学来确定蛋白质动态.
  • 该方法允许对H/D汇率进行定量评估.
  • 该研究概述了实施这种技术的实际步骤.

结论:

  • 提出的FT-IR光谱学协议为研究蛋白质动态提供了一个强大的方法.
  • 这种技术适用于研究突变,金属离子和连接体相互作用对蛋白质H/D交换的影响.
  • 该协议有助于更深入地了解生物系统中的结构功能关系.