INFLUENCES OF MACROMOLECULAR CROWDING ON BIOCHEMICAL SYSTEMS

大分子拥挤对生化系统的影响

• 批准号: 4689922
• 负责人: S B ZIMMERMAN
• 金额: --
• 依托单位: NAT INST OF ARTHRITIS, DIABETES, DIGESTIVE & KIDNEY DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/4689922
• 关键词: DNA; DNA repair; bacterial DNA; bacteriophage T4; bacteriophage lambda; chemical binding; circular DNA; deoxyribonucleotides; enzyme substrate; ethylene glycols; glycogen; macromolecule; molecular biology; molecular genetics; oligonucleotides; polymers; virus DNA

In our earlier studies of the effects of macromolecular crowding, we noted that the joining between DNA molecules catalyzed by T4 DNA ligase could be enormously stimulated by high concentrations of polymers. We have compared such crowding effects with those on the nonenzymatic cohesion between DNA molecules using assay systems based on the cohesive termini of Lambda DNA. High concentrations of several polymers greatly increase the rate of nonenzymatic cohesion, thereby allowing a test of the mechanism of the crowding effect on the DNA ligase reaction. The effects of macromolecular crowding were also examined with a different type of enzyme, T4 RNA ligase. Stimulation in the rates of joining of RNA substrates and the production of novel linear aggregates of RNA were obtained. Interactions with the Patent Office are proceeding in connection with the application on behalf of the Government of a patent covering the polymer-stimulated ligation of DNA.

在我们早期对大分子拥挤效应的研究中， 由T4 DNA连接酶催化的DNA分子之间的连接可能是 受到高浓度聚合物的极大刺激。 我们比较了 这种拥挤效应与DNA之间的非酶凝聚力 使用基于λ DNA的粘性末端的测定系统测定分子。 高浓度的几种聚合物大大增加了聚合物的速率。 非酶的凝聚力，从而允许测试的机制， DNA连接酶反应的拥挤效应。 大分子拥挤的影响也用不同的 T4 RNA连接酶 刺激RNA的连接速率 底物和RNA的新型线性聚集体的产生， 得到了 与专利局的互动正在进行中， 代表政府申请专利， 聚合物刺激的DNA连接。