对识别翻译后修改的质子"尾巴"的分析蛋白的方法
1Laboratory of Chemistry and Cell Biology, the Rockefeller University, New York, New York 10065, USA.
Journal of the American Chemical Society
|February 10, 2010
概括
研究人员开发了一种新型的探测器,以识别与修改后的组织蛋白相互作用的蛋白质. 这种工具专门捕获识别 histone H3 lysine-4三甲基化 (H3K4me3) 的蛋白质,有助于理解细胞过程.
科学领域:
- 表观遗传学和分子生物学
- 蛋白质组学和化学生物学
背景情况:
- 基因组后翻译修饰 (PTMs) 调节重要的细胞过程,如转录和染色体分离.
- 由于它们的动态和组合性质,对与基质子PTM相互作用的蛋白质进行全面的分析是具有挑战性的.
研究的目的:
- 开发一种新的化学生物学方法,用于识别直接与修饰的组织蛋白结合的蛋白质.
- 为了创建一个多功能探针来捕获和检测质子PTM结合蛋白.
主要方法:
- 设计了一种由蛋白质结构指导的光交叉链接探针,以共价连接相互作用的蛋白质.
- 包含一个基因组,用于点击化学介导的记者标签的结合 (用于检测的罗达胺,用于丰富的生物素).
- 开发并验证了在lysine-4 (H3K4me3) 中三甲基化素H3的特定探针.
主要成果:
- 成功设计和验证了一种能够捕获识别H3K4me3.3.的蛋白质的探针.
- 该探测器可以从整个蛋白质组中敏感检测和丰富目标蛋白的亲和力.
- 证明了探针用于识别特定的质子 PTM 相互作用蛋白的实用性.
结论:
- 这种新的探针设计可方便对与特定基因组PTMs相关的蛋白质进行全面的分析.
- 了解H3K4me3结合蛋白将为通过这种表观遗传标记调节的细胞功能提供新的见解.
- 开发的化学生物学策略广泛适用于研究其他质子PTM及其相互作用体.
相关概念视频
02:32Histone Modification
The histone proteins have a flexible N-terminal tail extending out from the nucleosome. These histone tails are often subjected to post-translational modifications such as acetylation, methylation, phosphorylation, and ubiquitination. Particular combinations of these modifications form “histone codes” that influence the chromatin folding and tissue-specific gene expression.
Acetylation
The enzyme histone acetyltransferase adds acetyl group to the histones. Another enzyme, histone deacetylase,...
Acetylation
The enzyme histone acetyltransferase adds acetyl group to the histones. Another enzyme, histone deacetylase,...
02:32Histone Modification
The histone proteins have a flexible N-terminal tail extending out from the nucleosome. These histone tails are often subjected to post-translational modifications such as acetylation, methylation, phosphorylation, and ubiquitination. Particular combinations of these modifications form “histone codes” that influence the chromatin folding and tissue-specific gene expression.
Acetylation
The enzyme histone acetyltransferase adds acetyl group to the histones. Another enzyme, histone deacetylase,...
Acetylation
The enzyme histone acetyltransferase adds acetyl group to the histones. Another enzyme, histone deacetylase,...
02:04Covalently Linked Protein Regulators
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.
These groups modify specific amino acids in a protein.
01:12The Nucleosome Core Particle
Nucleosomes are the DNA-histone complex, where the DNA strand is wound around the histone core. The histone core is an octamer containing two copies of H2A, H2B, H3, and H4 histone proteins.
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their primary aim is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. On the other hand, they must allow polymerase enzymes to access histone-bound DNA during...
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their primary aim is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. On the other hand, they must allow polymerase enzymes to access histone-bound DNA during...
02:25Spreading of Chromatin Modifications
The histone proteins in the nucleosomes are post-translationally modified (PTM) to increase or decrease access to DNA. The commonly observed PTMs are methylation, acetylation, phosphorylation, and ubiquitination of lysine amino acids in the histone H3 tail region. These histone modifications have specific meaning for the cell. Hence, they are called "histone code". The protein complex involved in histone modification is termed as "reader-writer" complex.
Writers
The writer is an enzyme that can...
Writers
The writer is an enzyme that can...