相关实验视频
Updated: Jun 17, 2026

12:07
Profiling Thiol Redox Proteome Using Isotope Tagging Mass Spectrometry
Published on: March 24, 2012
16.2K
使用无标签的化学蛋白质组学物质,对氨酸反应片段进行强大的蛋白质组概况
George S Biggs1,2,3, Emma E Cawood1,4, Aini Vuorinen1,2,3
1Crick-GSK Biomedical LinkLabs, GSK, Gunnels Wood Road, Stevenage, Hertfordshire, UK.
Nature communications
|January 2, 2025
概括
这项研究引入了一个新的无标签蛋白质组学平台,以快速识别药物点. 高通量方法有效地选氨酸反应碎片,加速发现人类蛋白质和潜在治疗药物的化学探针.
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
- 药物发现 药物发现 药物发现
背景情况:
- 鉴定人类蛋白质的药理探针对于治疗开发至关重要.
- 使用反应碎片的化学蛋白质组学是一种强大的联体发现方法,但在吞吐量,深度和可重现性方面面临挑战.
- 扩大可结合蛋白质组需要有效的方法来发现新的化学探针.
研究的目的:
- 开发和验证一个多功能,无标签的量化蛋白质组学平台,用于对氨酸反应片段的高通量选.
- 通过对本地蛋白质组进行对联体蛋白相互作用的分析来增强化学探针的发现.
- 解决样本吞吐量,蛋白质组深度和化学蛋白质组学中的数据可重复性方面的挑战.
主要方法:
- 使用了高通量,无标签量化蛋白质组学平台,结合了基于SP4板的样本准备和快速色谱渐变.
- 在 Bruker timsTOF Pro 2 上使用数据独立获取,用于对氨酸反应片段的竞争性分析.
- 选了80个对HEK293T和Jurkat细胞溶解物的反应碎片.
主要成果:
- 在每次运行中,我们始终确定了大约23,000个囊位,在两个细胞系中总共分析了约32,000个独特的囊位.
- 实现了高数据完整性,使联蛋白和 >400个联蛋白相互作用的稳定识别成为可能.
- 通过度-反应实验验验证了命中,并在命中扩张和活细胞实验中证明了实用性.
结论:
- 开发的无标签平台显著推进了高通量蛋白质组学,用于评估人类蛋白质组中氨酸结合性.
- 这种方法提供了一种多功能和可重复的方法,以加快药理学探针和潜在治疗方法的发现.
- 该平台能够有效地选和验证活性碎片,扩大化学生物学研究工具包.
相关概念视频
Protein Organization
Overview
Antibody Structure
Overview
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
Protein Organization
Proteins are polymers of amino acid residues. They are versatile and responsible for different cellular functions, including DNA replication, molecular transport, catalysis, and structural support. Proteins have a hierarchical structure comprising at least three levels of organization: primary, secondary, and tertiary structure. Some large proteins have a quaternary structure where individual protein subunits are linked together.
The primary structure of a protein is its amino acid sequence.
The primary structure of a protein is its amino acid sequence.
Tagging and Fusion Proteins
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
Western Blotting
Western blotting is an analytical technique for protein identification. It has various applications in immunology and medicine, including detecting diseases like bovine spongiform encephalopathy, mad cow disease, and human and feline immunodeficiency virus from biological samples.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.
Protein Organization
Proteins are polymers of amino acid residues. They are versatile and responsible for different cellular functions, including DNA replication, molecular transport, catalysis, and structural support. Proteins have a hierarchical structure comprising at least three levels of organization: primary, secondary, and tertiary structure. Some large proteins have a quaternary structure where individual protein subunits are linked together.
The primary structure of a protein is its amino acid sequence.
The primary structure of a protein is its amino acid sequence.

