通过DNA结合的聚胺 - 坎普托他辛合物对托酶I进行序列特定的捕获
1Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91125, USA.
Journal of the American Chemical Society
|September 6, 2001
概括
针对DNA的合成聚胺与camptothecin结合在一起. 这些结合物捕获Topoisomerase I,在特定地点诱导DNA裂变,结合物3显示出高效率.
科学领域:
- 分子生物学分子生物学
- 药用化学 医学化学
- 生物化学 生物化学
背景情况:
- role-imidazole聚胺是模仿DNA结合蛋白的合成配体.
- 这些分子与DNA的小沟结合,具有高度的亲和力和特异性.
研究的目的:
- 为了合成聚胺 - 坎普托他联体,以捕获Topoisomerase I.
- 使用这些结合物,在预先确定的部位诱导向的DNA裂变.
主要方法:
- 合成三种聚胺 - 坎普托他联体 (1-3) 具有不同的链接长度 (7,13,和18个原子).
- 在试验室中检测结合物捕获Topoisomerase I并诱导DNA裂变的能力.
- 确定由结合物诱导的DNA裂变的效率和特异性 3.
主要成果:
- 聚胺 - - 坎普托塞辛结合物已经成功合成.
- 结合物3在纳米分子度 (50nM) 上有效地捕获了Topoisomerase I.
- 结合物3诱导高产率 (77%) 的DNA从结合点在37°C时分离3-4个基对.
结论:
- 聚胺 - - 坎普托塞辛结合物是有针对性的DNA裂变的有效工具.
- 结合体3显示出对精确操纵DNA拓学的巨大潜力.
- 这种方法为控制酶介导的DNA相互作用提供了一个新的策略.
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...
DNA Damage can Stall the Cell Cycle
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...
Allosteric Proteins-ATCase
Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis pathway,...
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...
DNA Damage Can Stall the Cell Cycle
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...
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Bacterial pathogens depend on precise and efficient DNA replication to sustain infection. Two type II topoisomerases—DNA gyrase and topoisomerase IV—are critical to this process, as they resolve DNA supercoiling and unlink chromosomes during replication. Fluoroquinolones, synthetic derivatives of quinolones, exploit this mechanism by stabilizing the transient DNA–enzyme cleavage complex, preventing strand religation, and causing lethal double-strand breaks. These antibiotics are selectively...


