在接口处可逆DNA / 连接体复合物的融化热力学
Irina Belozerova1, Rastislav Levicky
1Department of Chemical & Biomolecular Engineering, Polytechnic Institute of New York University, Brooklyn, 11201, United States.
Journal of the American Chemical Society
|October 11, 2012
概括
本研究提出了一种基于表面的方法来分析DNA - 连接体相互作用,为溶液分析提供了一个可扩展和经济的替代方案. 该技术使用融过渡准确量化相互作用,与已建立的方法进行验证.
科学领域:
- 生物化学 生物化学
- 分子生物学分子生物学
- 分析化学 分析化学
背景情况:
- 分析小分子-核酸相互作用至关重要,但在规模上具有挑战性.
- 表面测定提供多重复合潜力,但在固体-液体界面面临量化障碍.
研究的目的:
- 调整一种基于溶液的方法,用于在表面上对DNA/配体相互作用的热力学分析.
- 为了将表面测定结果与溶液热量计数据进行比较.
- 建立基因与配体相互作用的基于表面的分析指南.
主要方法:
- 开发了一种表面测定方法,适应一种使用融过渡的溶液方法来量化DNA/连接物结合.
- 利用表面融过渡的电化学追踪进行热力学分析.
- 与使用网素作为模型连接体的溶液热量计数据对比的基准结果.
主要成果:
- 与溶液值相比,在DNA/网素相互作用的自由能量方面取得了很好的一致性.
- 通过可逆变化/杂化过渡确认了平衡操作.
- 证明了在表面上对DNA-配体相互作用的热力学分析的可行性.
结论:
- 开发的表面方法为DNA-连接体相互作用分析提供了一种可行,可扩展的方法.
- 制定了利用可逆化变性在表面的关键准则.
- 为了准确地分离体和体贡献,可能需要进一步的细化.
相关概念视频
DNA Topoisomerases
Topoisomerases are enzymes that relax overwound DNA molecules during various cell processes, including DNA replication and transcription. These enzymes regulate positive and negative DNA supercoiling without changing the nucleotide sequence. DNA overwinding in a clockwise direction results in positively supercoiled DNA, whereas underwinding in a counterclockwise direction produces negatively supercoiled DNA.
Types and Mechanism of action
Topoisomerases are divided into two main types. Type I...
Types and Mechanism of action
Topoisomerases are divided into two main types. Type I...
Phase Transitions: Melting and Freezing
Heating a crystalline solid increases the average energy of its atoms, molecules, or ions, and the solid gets hotter. At some point, the added energy becomes large enough to partially overcome the forces holding the molecules or ions of the solid in their fixed positions, and the solid begins the process of transitioning to the liquid state or melting. At this point, the temperature of the solid stops rising, despite the continual input of heat, and it remains constant until all of the solid is...
Protein-protein Interfaces
Many proteins form complexes to carry out their functions, making protein-protein interactions (PPIs) essential for an organism's survival. Most PPIs are stabilized by numerous weak noncovalent chemical forces. The physical shape of the interfaces determines the way two proteins interact. Many globular proteins have closely-matching shapes on their surfaces, which form a large number of weak bonds. Additionally, many PPIs occur between two helices or between a surface cleft and a polypeptide...
Ligand Binding and Linkage
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 the...
Single-Strand DNA Binding Proteins
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...
Ligand Binding Sites
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...


