拓糖酶II N端ATPase粘合剂 稳定性 粘合剂
Lauren A Fielding1, Joseph Deweese2,3
1Department of Biological, Physical, and Human Sciences, Freed-Hardeman University, Henderson, TN, USA.
Methods in molecular biology (Clifton, N.J.)
|May 15, 2025
概括
这项研究引入了一种拓酶II试验,以测量酶-DNA复合物的稳定性. 该方法量化了不同条件如何影响拓酶-DNA相互作用的强度.
科学领域:
- 生物化学 生化学
- 分子生物学分子生物学
- 酶学 是一种酶学.
背景情况:
- 拓二酶酶对DNA复制和修复至关重要.
- 了解拓酶-DNA相互作用的稳定性是理解酶功能的关键.
研究的目的:
- 开发和验证一种新的测定方法,用于评估多酶II-DNA复合物的稳定性.
- 调查影响N端ATPase紧剂与DNA相互作用稳定性的因素.
主要方法:
- 使用了一种拓聚酶II N-终端ATPase稳定试验.
- 蛋白质:DNA复合体被固定并受到越来越严格的冲洗.
- 在每个洗步骤中解的DNA通过凝电泳量化.
主要成果:
- 该试验有效地确定了拓酶II-DNA相互作用的相对稳定性.
- 洗的严格性与酶:DNA复合物的破坏相关.
- 该方法允许识别稳定这些复合物的条件.
结论:
- 开发的试验为研究酶-DNA复杂动力学提供了一个强大的方法.
- 这种技术有助于探索调控托波酶II活性的调节机制.
- 这些发现有助于更深入地了解由topoisomerase II进行的DNA拓管理.
更多相关视频
10:12Author Spotlight: Quantitative Detection of DNA Protein Crosslinks and Their Post-Translational Modifications
Published on: April 21, 2023
2.6K
12:19Tools to Study the Role of Architectural Protein HMGB1 in the Processing of Helix Distorting, Site-specific DNA Interstrand Crosslinks
Published on: November 10, 2016
8.2K
相关概念视频
DNA Topoisomerases
30.6K
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. ...
Types and Mechanism of action
Topoisomerases are divided into two main types. ...
30.6K
Destabilization of Microtubules
2.5K
The destabilization of microtubules can occur during different stages of the microtubule lifecycle, such as nucleation or elongation. It can take place at either end of the microtubule or in the microtubule lattices as a whole. The lifespan of individual microtubules within a cell varies according to the cell type and stage of the cell cycle. During interphase, the lifespan of the microtubule is about 30 minutes, while during cell division, it is about 15 minutes. In axonal microtubules of...
2.5K
Restarting Stalled Replication Forks
5.7K
DNA replication is initiated at sites containing predefined DNA sequences known as origins of replication. DNA is unwound at these sites by the minichromosome maintenance (MCM) helicase and other factors such as Cdc45 and the associated GINS complex.The unwound single strands are protected by replication protein A (RPA) until DNA polymerase starts synthesizing DNA at the 5’ end of the strand in the same direction as the replication fork. To prevent the replication fork from falling apart,...
5.7K
The Replisome
32.7K
DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
32.7K
Microtubule Instability
4.9K
Microtubules are hollow cylindrical filaments having a diameter of approximately 25 nm and a length that varies from 200 nm to 25 μm. GTP-bound tubulin subunits form αβ-heterodimers for microtubule assembly. These core building blocks interact longitudinally, polymerizing into protofilaments. The protofilaments then interact with one another through lateral bonding forces to form stable cylindrical microtubules. These cylindrical filaments are dynamic as they undergo repeated...
4.9K
DNA Damage can Stall the Cell Cycle
9.0K
In response to DNA damage, cells can pause the cell cycle to assess and repair the breaks. However, the cell must check the DNA at certain critical stages during the cell cycle. If the cell cycle pauses before DNA replication, the cells will contain twice the amount of DNA. On the other hand, if cells arrest after DNA replication but before mitosis, they will contain four times the normal amount of DNA. With a host of specialized proteins at their disposal,cells must use the right protein at...
9.0K
