将MicroED和原生质谱相结合,用于酶-小分子复合体的结构发现
Niko W Vlahakis1,2,3, Cameron W Flowers1,2,3, Mengting Liu4
1Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095.
概括
我们使用原生质谱 (ED-MS) 开发了电子衍射,以快速发现小分子-蛋白质复合体. 这种方法通过分析来自原始样本的蛋白质-配体相互作用来加速基于结构的药物发现.
科学领域:
- 结构生物学 结构生物学
- 生物化学 生物化学
- 药物发现 药物发现 药物发现
背景情况:
- 加速发现小分子-蛋白质复合体对于药物开发至关重要.
- 目前用于结构确定的方法可能耗时,需要净化样本.
研究的目的:
- 开发和验证一种新的方法,即使用原生质谱 (ED-MS) 进行电子衍射,用于快速识别和结构性描述小分子-蛋白质复合体.
- 证明ED-MS在发现天然产品配体和选小分子结合物的有用性.
主要方法:
- 使用快速,低剂量,基于事件的电子计数微晶电子衍射 (MicroED) 数据收集.
- 集成的MicroED与本地质谱仪 (MS).
- 将ED-MS应用于已知抑制剂浸泡的晶体泥,原始生物合成反应和打印的配体库.
主要成果:
- 成功地解决了E-64及其与papain结合的类似物的结构.
- 确定了与帕帕因结合的首选天然产品,并检测了来自未经净化原始生物合成反应的复合物.
- 确定了阿维巴克坦与CTX-M-14β-乳糖酶的结合,包括化合物混合物中的结合.
结论:
- ED-MS是一种强大的技术,可以加速发现小分子-蛋白质复合体.
- 该方法可以使用原始样本对依干与宏分子标进行基于结构的选.
- 在推进药物发现工作方面,ED-MS显示出显著的前景.
相关概念视频
Mass Spectrometry: Complex Analysis
897
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...
897
Peptide Identification Using Tandem Mass Spectrometry
6.8K
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...
6.8K
Mass Spectrometry: Overview
5.9K
Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass. One common type of ionization, known as electrospray ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave...
5.9K
Proteomics
7.9K
A proteome is the entire set of proteins that a cell type produces. We can study proteomes using the knowledge of genomes because genes code for mRNAs, and the mRNAs encode proteins. Although mRNA analysis is a step in the right direction, not all mRNAs are translated into proteins.
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...
Proteomics is the study of proteomes' function. It involves the large-scale systematic study of the proteome to denote the protein complement expressed by a genome. Scientist Mark Wilkins coined the term...
7.9K


