翻訳後に改変されたヒストンの"尾"を認識するプロファイルタンパク質へのアプローチ
1Laboratory of Chemistry and Cell Biology, the Rockefeller University, New York, New York 10065, USA.
Journal of the American Chemical Society
|February 10, 2010
まとめ
研究者は,改変されたヒストンと相互作用するタンパク質を識別するための新しい探査機を開発しました. このツールは,ヒストンH3ライシン-4トリメチル化 (H3K4me3) を認識するタンパク質を特に捕捉し,細胞のプロセスを理解するのに役立ちます.
科学分野:
- エピジェネティクスと分子生物学
- プロテオミクスと化学生物学
背景:
- ヒストンの翻訳後の改変 (PTMs) は,転写や染色体分離のような重要な細胞過程を調節する.
- ヒストンPTMと相互作用するタンパク質の包括的なプロファイリングは,それらのダイナミックで組合せ的な性質のために困難です.
研究 の 目的:
- 改変ヒストンに直接結合するタンパク質を特定するための新しい化学生物学アプローチを開発する.
- ヒストンPTM結合タンパク質を捕捉および検出するための多用途のプローブを作成する.
主な方法:
- 相互作用するタンパク質を共振的に結合するために,タンパク質構造によって導かれるフォトクロスリンカープローブを設計した.
- クリック化学によるレポータータグの結合 (検出のためのロダミン,濃縮のためのバイオチン) のためのアルキン群を組み込みました.
- リジン-4 (H3K4me3) でトリメチル化ヒストンH3の特定のプローブを開発し,検証しました.
主要な成果:
- H3K4me3.3.を認識するタンパク質を捕捉できる探査機を成功裏に設計・検証しました.
- 探査機は,タンパク質全体から標的タンパク質の感度検知と親和濃縮を可能にします.
- 特定のヒストロン PTM 相互作用タンパク質を特定するための探査機の有用性を実証しました.
結論:
- この新しい探査装置の設計は,特定のヒストンPTMに関連したタンパク質の包括的なプロファイリングを容易にする.
- H3K4me3結合タンパク質の理解は,この表遺伝子マークによって調節される細胞機能に関する新しい洞察を提供します.
- 開発された化学生物学戦略は,他のヒストンPTMとその相互作用体を研究するのに広く適用できます.
関連する概念動画
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...