相关实验视频
Updated: Jun 23, 2026
09:47
Analysis of Histone Antibody Specificity with Peptide Microarrays
Published on: August 1, 2017
基因组 H4-K16 乙特异性 DNA 胺体的演化
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
概括
我们开发了DNA体,专门结合乙化组合素H4. 这些体为翻译后修改提供了高特异性,在选择性方面超过了抗体.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 表观遗传学 在表观遗传学中,表观遗传学是指表观遗传学.
背景情况:
- 基因组转化后修饰 (PTMs) 调节基因表达.
- 在 lysine 16 (H4K16ac) 中的 histone H4 乙化是一个关键的表观遗传标记.
- 开发用于检测基因组PTMs的特定工具对于理解表观遗传调节至关重要.
研究的目的:
- 为了生成特定识别和结合在lysine 16 (H4K16ac) 处乙化的基因组 H4 的DNA 体.
- 为了描述所选的体的结合亲和力和特异性.
- 为了比较aptamers与用于检测H4K16ac的商业抗体的性能.
主要方法:
- 在体外选择 (SELEX) 以确定向H4K16ac.的DNA受体.
- 表面等离子体共振 (SPR) 用于确定结合亲和力 (K(d)).
- 竞争性结合试验用于评估对非乙化素H4和在lysine 8 (H4K8ac) 中乙化H4的特异性.
主要成果:
- 为H4K16ac.选择了一种具有高亲和度 (K(d) = 21nM) 的DNA吸收体.
- 胺体表现出显著的特异性,区分非乙化 histone H4 和 H4K8ac.
- 与商业抗体相比,阿普坦酶表现出类似的结合亲和力,但特异性更高 (比2400倍高15倍).
结论:
- 可以生成DNA体,专门针对蛋白质的翻译后修改.
- 亚胺可以实现对修饰及其在蛋白质上的位置的高特异性.
- 这些体体为表观遗传研究和诊断提供了宝贵的工具.
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,...
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...