对于高含量的化学蛋白质组学而言,一个双向直角蛋白质分解策略
Anna E Speers1, Benjamin F Cravatt
1The Skaggs Institute for Chemical Biology and The Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA.
Journal of the American Chemical Society
|July 14, 2005
概括
这项研究介绍了一种高含量化学蛋白质组的双向直角蛋白质分解 (TOP) 策略. 这种方法有效地识别了基于活动的蛋白质分析 (ABPP) 探针的蛋白质标和修改点.
科学领域:
- 蛋白质组学是指蛋白质组学.
- 化学生物学 化学生物学
- 酶学 是一种酶学.
背景情况:
- 蛋白质组学旨在功能性地注释基因组,使用基于活动的蛋白质概况 (ABPP) 等化学策略,通过功能来丰富蛋白质.
- 现有的分析技术限制了化学蛋白质原子实验中获得的数据.
- 需要先进的方法来表征探针标记蛋白质及其修改位点.
研究的目的:
- 开发一种高含量化学蛋白质组学策略,用于对探头标记蛋白质和修饰位点进行并行表征.
- 为了应用这一策略来识别硫酸盐的标,ABPP在组织蛋白质组中的探针.
主要方法:
- 开发一个双向直角蛋白质分解 (TOP) 战略.
- 使用"点击化学"用生物素和烟草蚀刻病毒 (TEV) 蛋白质分裂部位标记探针标记的蛋白质.
- 蛋白质和标记的序列丰富和识别,使用斯特雷普塔维丁珠和两步蛋白质分解 (素和TEV).
主要成果:
- 在组织蛋白质组中成功应用TOP方法来表征酸盐ABPP探针的标.
- 发现了许多活性位点标记酶.
- 在调节部位上修改的酶和功能不明的蛋白质的识别.
结论:
- TOP策略使高含量化学蛋白质学成为可能,允许对探测器目标和修改站点进行并行表征.
- 硫酸盐探针针对各种各样的功能残留物,包括活性和调节部位.
- 基于TOP的化学蛋白质学对于表征蛋白质及其调节残留物非常有价值.
相关概念视频
01:33Peptide Identification Using Tandem Mass Spectrometry
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...
01:33Proteomics
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...
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...
01:21Tandem 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...