使用可变温度离子运动质谱法探索刚性和灵活分子的 conformational 景观
Xudong Wang1, Emma Norgate1, Junxiao Dai1
1Michael Barber Centre for Collaborative Mass Spectrometry, Manchester Institute of Biotechnology, Department of Chemistry, The University of Manchester, 131 Princess Street, Manchester, M1 7DN, UK.
Nature communications
|May 5, 2025
概括
可变温度离子移动性质谱法 (VT-IM-MS) 揭示了温度如何影响蛋白质结构. 该方法将结构变化与碰撞区分开来,为蛋白质热力学提供了洞察力.
科学领域:
- 生物物理化学 生物物理化学
- 结构生物学是结构生物学.
- 质谱测量质量谱测量
背景情况:
- 蛋白质的结构完整性在生物技术和气候变化研究中至关重要.
- 可变温度离子流动性质谱法 (VT-IM-MS) 是一种研究温度依赖的蛋白质构造变化的技术.
研究的目的:
- 为了研究温度对蛋白质结构完整性和形状景观的影响.
- 使用VT-IM-MS.区分碰撞效应与实际结构变化.
主要方法:
- 在190350K的温度范围内对聚 (L-氨酸) (PLL) 树枝状物,无素,β-氨酸和α-氨酸进行了测量.
- 使用可变温度离子移动性质谱法 (VT-IM-MS) 来分析形状变化.
- 碰撞诱导解离 (CID) 效应被认为与结构变化有所区别.
主要成果:
- PLL树枝状体的碰撞横截面 (CCS) 显示了与碰撞理论相一致的温度依赖.
- 蛋白质 (ubiquitin,β-casein,α-synuclein) 在350K和250K显著重组,与体外稳定性预测保持一致.
- 展开的中间体在较低的温度 (210 K和190 K) 时被动态捕获.
- 对于α-synuclein,观察到不同的适配体,使得可以计算过渡速率和激活能.
结论:
- VT-IM-MS有效地区分了温度诱导的结构变化和碰撞效应.
- 这项研究表明,VT-IM-MS能够为蛋白质结构重组提供热力学见解.
- 动力捕获的中间体为蛋白质展开的途径提供了一个窗口.
相关概念视频
¹H NMR of Conformationally Flexible Molecules: Variable-Temperature NMR
1.0K
The axial and equatorial protons in cyclohexane can be distinguished by performing a variable-temperature NMR experiment. In this process, except for one proton, the remaining eleven protons are replaced by deuterium. The deuterium substitution avoids the possible peak splitting caused by the spin-spin coupling between the adjacent protons. The remaining proton flips between the axial and equatorial positions.
1.0K
¹H NMR of Conformationally Flexible Molecules: Temporal Resolution
780
At room temperature, the chair conformer of cyclohexane undergoes rapid ring flipping between two equivalent chair conformers at a rate of approximately 105 times per second. These two chair conformers are in equilibrium. The rapid ring flipping results in the interconversion of the axial proton to an equatorial proton and an equatorial to the axial proton. Such interconversions are too rapid and cannot be detected on the NMR timescale. Hence, the NMR spectrometer cannot distinguish between the...
780
High-Resolution Mass Spectrometry (HRMS)
1.1K
The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For...
1.1K
Mass Spectrometry: Complex Analysis
652
Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
652
Electrospray Ionization (ESI) Mass Spectrometry
664
Higher molecular weight biomolecules are nonvolatile compounds that may decompose before ionizing or vaporizing during mass analysis with conventional electron impact ionization methods. Accordingly, electrospray ionization (ESI) is the favored method for vaporizing and ionizing biomolecules as it circumvents rapid fragmentation and enables the recording of mass signals for the entire biomolecule.
ESI utilizes electrical energy to transfer ions from the liquid phase of the sample into the...
ESI utilizes electrical energy to transfer ions from the liquid phase of the sample into the...
664
Peptide Identification Using Tandem Mass Spectrometry
6.2K
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
6.2K


