-交换质谱识别了多元组分激素SAM酶,PqqE内的局部和远距离相互作用
Wen Zhu1,2, Anthony T Iavarone2, Judith P Klinman2,3,4
1Department of Chemistry and Biochemistry, Florida State University, Tallahassee, Florida 32306, United States.
ACS central science
|March 4, 2024
概括
-交换质谱法 (HDX-MS) 揭示了类 (PQQ) 生物合成中的蛋白质动态. 这项研究描述了对PqqE酶功能和天然产品合成至关重要的蛋白质-相互作用.
科学领域:
- 生物化学 生物化学
- 结构生物学 结构生物学
- 酶学 是一种酶学.
背景情况:
- 蛋白质-相互作用对生物过程至关重要,包括天然产品生物合成.
- 激进的SAM (rSAM) 酶PqqE启动了从前体中产生类 (PQQ) 的生产.
研究的目的:
- 用HDX-MS.来描述参与PqqE功能中的蛋白质-蛋白质和蛋白质-化合物的结构动态.
- 为了研究基质和辅因子与PqqE/PqqD复合体结合的结构变化.
主要方法:
- -交换质谱法 (HDX-MS) 用于绘制蛋白质-相互作用和构造动态.
- 尺寸排除色谱与小角度X射线散射 (SEC-SAXS) 结合,以支持结构发现.
主要成果:
- 在PqqA,PqqD和PqqE的二元和三元复合体中,HDX-MS确定了关键的相互作用区域 ("热点").
- 在基质 (PqqA) 和S-adenosylmethionine (SAM) 与PqqE/PqqD复合体结合时观察到相互的构造变化.
- 在四级复合体中检测到大约40 Å的远程形状变化,将PqqA结合部位与PqqE活性部位连接起来.
结论:
- HDX-MS有效地表征了蛋白质构造通信在改 rSAM 酶中的特征.
- 直接接触和远距离通信都在PqqE功能复合体内的相互作用中介.
- 该方法适用于研究涉及基修饰的广泛rSAM酶.
更多相关视频
09:18Time-resolved ElectroSpray Ionization Hydrogen-deuterium Exchange Mass Spectrometry for Studying Protein Structure and Dynamics
Published on: April 17, 2017
9.9K
11:32A Hydrogen-Deuterium Exchange Mass Spectrometry HDX-MS Platform for Investigating Peptide Biosynthetic Enzymes
Published on: May 4, 2020
8.0K
相关概念视频
01:17Electron Paramagnetic Resonance (EPR) Spectroscopy: Organic Radicals
2.5K
Ideally, an unpaired electron shows a single peak in the EPR spectrum due to the transition between the two spin energy states. However, coupling interactions can occur between the spins of the unpaired electron and any neighboring spin-active nuclei. This hyperfine coupling results in hyperfine splitting, where the EPR signal is split into multiplets. The signals split into 2nI + 1 peaks, where n is the number of equivalent nuclei and I is the nuclear spin. These splitting patterns provide...
2.5K
00:48¹H NMR of Labile Protons: Deuterium (²H) Substitution
886
This lesson illustrates the role of deuterium substitution in simplifying the NMR spectrum of compounds comprising labile protons. One method employed is the use of deuterium. Amongst the three isotopes of hydrogen, deuterium (2H) has a nucleus composed of one proton and one neutron. When the D2O solvent is added to a pure dry ethanol solution, its labile proton is substituted with deuterium.
886
01:11Radical Reactivity: Overview
2.1K
Radicals, the highly reactive species, gain stability by undergoing three different reactions. The first reaction involves a radical-radical coupling, in which a radical combines with another radical, forming a spin‐paired molecule. The second reaction is between a radical and a spin‐paired molecule, generating a new radical and a new spin‐paired molecule. The third reaction is radical decomposition in a unimolecular reaction, forming a new radical and a spin‐paired...
2.1K