関連する実験動画
Updated: Jun 23, 2026
09:47
Analysis of Histone Antibody Specificity with Peptide Microarrays
Published on: August 1, 2017
ヒストンH4-K16アセチル固有のDNAアプタメアの進化
Berea A R Williams1, Liyun Lin, Stuart M Lindsay
1Center for BioOptical Nanotechnology, Arizona State University, Tempe, Arizona 85287-5201, USA.
Journal of the American Chemical Society
|April 24, 2009
まとめ
私たちは,アセチル化ヒストンH4に特異的に結合するDNAアプタマーを開発しました. これらのアプタマーは,翻訳後の改変に対して高い特異性を提供し,選択性において抗体を上回ります.
科学分野:
- 分子生物学は分子生物学である.
- バイオケミストリー バイオケミストリー
- エピジェネティクス エピジェネティクス
背景:
- ヒストンの翻訳後の改変 (PTMs) は遺伝子発現を調節する.
- リジン16 (H4K16ac) でのヒストンH4アセチル化が重要な表遺伝的マーカーである.
- ヒストンPTMを検出するための特定のツールの開発は,表遺伝的調節を理解するために不可欠です.
研究 の 目的:
- リスイン16 (H4K16ac) でアセチル化されたヒストンH4に特異的に認識し,結合するDNAアプタマーを生成する.
- 選択されたアプタマーの結合親和性と特異性を特徴付ける.
- H4K16ac検出のための商用抗体に対するアプタマーの性能を比較するために.
主な方法:
- H4K16ac.を標的とするDNAアプタマーを特定するためのインビトロ選択 (SELEX)
- 表面プラズモン共振 (SPR) で結合親和度 (K(d)) を決定する.
- コンペティティブ・バインディング・アッセイは,ライシン8 (H4K8ac) でアセチル化された非アセチルヒストーンH4およびH4に対する特異性を評価する.
主要な成果:
- H4K16ac.ac.に対する高い親和性 (K(d) = 21 nM) を有するDNAアプタマーが選択されました.
- アプタミーは有意な特異性を示し,非アセチル化ヒストーンH4およびH4K8ac.ac.を区別しました.
- 商用抗体と比較すると,アプタメルは類似の結合親和性を示したが,特異性はかなり高かった (15倍対2400倍).
結論:
- DNAアプタマーは,特異的にタンパク質の翻訳後の修正をターゲットに生成することができます.
- アプタマーは,修正とタンパク質上の位置の両方に高い特異性を達成することができます.
- これらのアプタマーは,表遺伝子研究と診断のための貴重なツールです.
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,...
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...
02:10The 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.
The paradox
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their main responsibility is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. While on the other hand, they must allow polymerase enzymes to access DNA...
The paradox
Nucleosomes, paradoxically, perform two opposite functions simultaneously. On the one hand, their main responsibility is to protect the delicate DNA strands from physical damage and help achieve a higher compaction ratio. While on the other hand, they must allow polymerase enzymes to access DNA...
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...
02:30Histone Variants at the Centromere
Histone variants are the histone proteins with structural and sequence variations. These variants may be regarded as “mutant” forms that replace their canonical histone counterparts in the nucleosomes. Specific post-translational modifications on the histone variants enable further chromatin complexity and regulate tissue-specific gene expression. The most common histone variants are from histone H2A, H2B, and linker histone H1 families. However, several variants of histone H3 variants are also...