单链或双链DNA标签对DNA编码图书馆中的带结合亲缘关系的影响
Philipp Bittner1, Andreas Gloger2, Julian A Harrison1
1Laboratory of Organic Chemistry, Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland.
Analytical chemistry
|September 9, 2025
概括
用DNA编码的库对于药物发现至关重要. 这项研究表明,DNA标签结构显著影响化合物结合亲和力,单链DNA标签显示出协同结合效应.
科学领域:
- 生物化学 生化学
- 药物发现 药物发现 药物发现
- 分子生物学分子生物学
背景情况:
- DNA编码图书馆 (DEL) 是药物发现的强大工具.
- 存在各种DNA标签格式 (单链,双链) 和链接器化学.
- 之前的研究指出了DNA标签格式之间的结合亲和力差异,但分子基础尚不清楚.
研究的目的:
- 调查DNA标签对DELs蛋白质-配体结合性能的影响.
- 阐明结合亲缘关系观察到的差异的分子基础.
- 评估DNA标签结构对结合动态的影响.
主要方法:
- 利用原生离子流动性质谱法.
- 集成的气相和溶液相激活技术.
- 系统地分析了与不同DNA标签的蛋白质-连接体相互作用.
主要成果:
- 结合DNA合化合物的结合亲和力受到DNA标签设置的强烈影响.
- 结合 afinities 的差异与 On-DNA 蛋白质 - 连接体复合物的热稳定性相关.
- 在各种系统中,只观察到与单链DNA合的配体的协同结合效应.
结论:
- DNA标签格式对DELs的结合亲和力产生了重大影响.
- 质谱法可以阐明DNA标签如何影响结合动态.
- 了解这些影响对于优化DEL技术中的选和命中验证至关重要.
相关概念视频
Labeling DNA Probes
9.3K
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...
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...
9.3K
Single-Strand DNA Binding Proteins
16.6K
For successful DNA replication, the unwinding of double-stranded DNA must be accompanied by stabilization and protection of the separated single strands of the DNA. This crucial task is performed by single-strand DNA-binding (SSB) proteins. They bind to the DNA in a sequence-independent manner, which means that the nitrogenous bases of the DNA need not be present in a specific order for binding of SSB proteins to it. The binding of SSB proteins straightens single-stranded DNA (ssDNA) and makes...
16.6K
The Equilibrium Binding Constant and Binding Strength
14.9K
The equilibrium binding constant (Kb) quantifies the strength of a protein-ligand interaction. Kb can be calculated as follows when the reaction is at equilibrium:
14.9K
Tagging and Fusion Proteins
8.3K
Proteins are involved in several cellular processes and biochemical reactions. Analyzing a specific protein of interest requires it to be isolated from the other proteins in the cell. This is achieved by overexpressing the specific gene in a suitable host to produce large quantities of the target protein. A tag or label is recombined with the gene to produce a fusion protein containing the target protein and the tag. The tags on these fusion proteins can then be used for easy detection and...
8.3K
Ligand Binding Sites
14.9K
Proteins are dynamic macromolecules that carry out a wide variety of essential processes; however, the activities of most proteins depend on their interactions with other molecules or ions, known as ligands.
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
Protein-ligand interactions are quite specific; even though numerous potential ligands surround a cellular protein at any given time, only a particular ligand can bind to that protein. Moreover, a ligand binds only to a dedicated area on the surface of the protein, known as the...
14.9K
Ligand Binding and Linkage
5.5K
Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked. In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...
5.5K


