在大肠杆菌中特定地点的胺/交换电喷离子质谱法测量出的胺
1Department of Chemistry, Oregon State University, Corvallis, OR 97331, USA.
Journal of the American Chemical Society
|October 5, 2001
概括
使用b(n) 离子的质谱可靠地测量蛋白质的吸收,补充NMR研究蛋白质结构和折叠动态.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 结构生物学 结构生物学
背景情况:
- 与质谱学 (MS) 结合的/交换是蛋白质分析的新兴技术.
- 了解蛋白质折叠和结构在分子生物学中至关重要.
研究的目的:
- 为了评估质谱学 (MS) 的可靠性,使用b (n) 和y (n) 离子进行特定地点的胺含量测量.
- 通过/交换研究大肠杆菌硫的蛋白质折叠和结构特征.
主要方法:
- 测量了氧化和减少的大肠杆菌 thioredoxins. 在特定地点的胺基含量.
- 电子喷雾电离CIDMS/MS实验是在D(2) O中化后进行的.
- 来自b (n) 和y (n) 离子的数据与NMR确定的H/D汇率常数进行了比较.
主要成果:
- 通过b(n) 离子测量的含量与NMR数据相关性良好,准确地反映了区域蛋白质结构.
- 阿尔法螺旋和β片中的残留物显示出低合物,而循环和转显示出高合物.
- 在Cys-32的化诱导了轻微的结构变化,具体残留物中的含量略高表明了这一点.
结论:
- 质谱法,特别是使用b(n) 离子,是评估特定地点的胺同位素交换的可靠方法.
- 马氏素作为一种有价值的NMR补充技术,用于探测蛋白质结构特征.
更多相关视频
11:32A Hydrogen-Deuterium Exchange Mass Spectrometry (HDX-MS) Platform for Investigating Peptide Biosynthetic Enzymes
Published on: May 4, 2020
08:40Millisecond Hydrogen/Deuterium-Exchange Mass Spectrometry for the Study of Alpha-Synuclein Structural Dynamics Under Physiological Conditions
Published on: June 23, 2022
相关概念视频
¹³C NMR: Distortionless Enhancement by Polarization Transfer (DEPT)
When proton-coupled carbon-13 spectra are simplified by a broadband proton decoupling technique, structural information about the coupled protons is lost. Distortionless enhancement by polarization transfer (DEPT) is a technique that provides information on the number of hydrogens attached to each carbon in a molecule. While the DEPT experiment utilizes complex pulse sequences, the pulse delay and flip angle are specifically manipulated. The resulting signals have different phases depending on...
Tandem Mass Spectrometry
Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...
¹H NMR of Labile Protons: Deuterium (²H) Substitution
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.
Electrospray Ionization (ESI) Mass Spectrometry
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...
