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链接器 H1 基因组直接上下蛋白形状分析
Md Shofiul Alam1, Cassandra N Fuller1, Kevin Jeanne Dit Fouque1
1Department of Chemistry and Biochemistry, Florida International University, Miami, Florida 33199, United States.
Journal of proteome research
|September 23, 2025
概括
我们开发了一种新型的质谱法,可以直接鉴定链接组织素H1蛋白形及其后翻译修饰 (PTMs). 这种技术有效地识别H1变异及其PTM,推进染色体研究.
科学领域:
- 蛋白质组学是指蛋白质组学.
- 染色体生物学 染色体生物学
- 质谱测量质量谱测量
背景情况:
- 链接器H1基因素对染色质结构和基因调节至关重要.
- 翻译后修改 (PTMs) 显著影响H1质子功能.
- 对H1蛋白形及其PTM的直接表征仍然具有挑战性.
研究的目的:
- 开发和验证一种基于直接质谱的方法,用于表征链接器H1基因组蛋白质形式.
- 为了识别和映射H1基因组变体上的各种PTM.
- 为了使H1蛋白形异质性的全面分析.
主要方法:
- 捕获的离子移动性光谱与紫外线光解离和富里埃变换离子循环子质谱学相结合 (TIMS-q-UVPD-FT-ICR MS/MS).
- 在碎片化之前,基于移动性和质量的蛋白质形式的预制.
- 高质量精度检测碎片离子用于PTM分配.
- 补充自上而下的 (LC-q-EAD-ToF MS/MS) 和自下而上的分析用于验证.
主要成果:
- 四种牛H1变种 (H1.2,H1.3,H1.5,H1.4V) 及其PTM的直接表征.
- 对于已识别的H1蛋白形来说,实现了高序列覆盖率 (高达60%).
- 识别各种PTM,包括单/二甲基化,乙化和酸化.
- 新的测序成功识别了H1.4V变体的序列.
结论:
- 该TIMS-q-UVPD-FT-ICRMS/MS方法提供了H1蛋白形及其PTM的直接和有效的表征.
- 这种方法需要最小的样本准备,并提供了广泛的PTM观测.
- 该方法对染色体研究中的全球H1蛋白形分析具有重大前景.
关键词:
这是自下而上的.染色体重塑 染色体重塑 的方法非洲发展署 (EAD) 已经开始.在FT-ICR MS中使用.链接器组织基因组.后翻译修改后的修改.蛋白质形式的蛋白质.这里是 TIMS TIMS 的时间.从上到下,从上到下.紫外线PD 紫外线PD 的情况.在 de novo 测序中使用 de novo 测序.更多相关视频
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相关概念视频
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.
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,...
02:30Histone 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...
02:38Heterochromatin
The extent of chromatin compaction can be studied by staining chromatin using specific DNA binding dyes. Under the microscope, the dense-compacted regions that take up more dye are called heterochromatin. Heterochromatin is further classified into two forms – constitutive heterochromatin and facultative heterochromatin.
Constitutive heterochromatin: It is a highly compact region of chromatin that is mostly concentrated in the centromere and telomere. Unlike euchromatin, the amino acid at 9th...
Constitutive heterochromatin: It is a highly compact region of chromatin that is mostly concentrated in the centromere and telomere. Unlike euchromatin, the amino acid at 9th...
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...