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Labeling DNA Probes

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Updated: Sep 10, 2025

Analysis of Histone Antibody Specificity with Peptide Microarrays

Analysis of Histone Antibody Specificity with Peptide Microarrays

Published on: August 1, 2017

通过乙化H2A.Z光敏度探针对BPTF目标进行接触

Kerstin E Peterson¹, Noelle M Olson¹, Dani J Dahlseid¹

¹Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, United States.

|August 27, 2025

概括

研究人员开发了新的光亲和探测器来研究乙化组合素H2A.Z和BPTF之间的相互作用,这是与癌症进展相关的蛋白质. 这些探测器捕获了BPTF与H2A.Z的结合,揭示了肺癌细胞中关键的乙化模式.

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Last Updated: Sep 10, 2025

Analysis of Histone Antibody Specificity with Peptide Microarrays

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Identification of Small Molecule-binding Proteins in a Native Cellular Environment by Live-cell Photoaffinity Labeling

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科学领域:

表观遗传学
癌症生物学
蛋白质组学

背景情况:

基因组变异H2A.Z与癌症的进展有关.
在癌症中,表观遗传阅读蛋白BPTF的功能往往失调.
由于结合亲缘关系较弱,乙化H2A.Z与BPTF之间的相互作用缺乏直接验证.

研究的目的:

开发和使用光亲和探测器来捕获乙化H2A.Z单体和BPTF之间的短暂相互作用.
验证BPTF和乙化H2A.Z之间的直接相互作用.
在人类非小细胞肺癌细胞中识别H2A.Z的天然乙化模式.

主要方法:

开发具有N终端迪亚齐林和C终端生物标记的光亲和性乙化素H2A.Z探针 (I和II异形).
通过SDS-PAGE进行相交链接实验以评估BPTF-H2A.Z的相互作用.
下向蛋白质组测量A549非小细胞肺癌细胞中的H2A.Z乙化模式.

主要成果:

复合BPTF原蛋白直接与两个H2A.Z异型相互作用,这种相互作用依赖于亲和力和乙-氨酸结合的特定方式.
内源BPTF通过A549细胞核溶解物中的乙化H2A.Z探针进行光交联,尽管效率低于H4K16ac.
蛋白质组学发现H2A.Z的单乙烯和二乙烯化是主要的模式,其中包括K4ac,K7ac,K11ac和K15ac的特定乙烯化位点.

结论:

光亲和探测器有效捕获涉及H2A.Z的过渡性表观蛋白相互作用.
这项研究证实了BPTF和乙化H2A.Z之间的直接相互作用.
需要进一步优化,通过互动组分析识别额外的H2A.Z表观遗传调节剂.