INFLUENCES OF MACROMOLECULAR CROWDING ON BIOCHEMICAL SYSTEMS

大分子拥挤对生化系统的影响

• 批准号: 3940872
• 负责人: S B ZIMMERMAN
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DIABETES AND DIGESTIVE AND 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/3940872
• 关键词: DNA; DNA directed DNA polymerase; bacterial DNA; bacteriophage T4; bacteriophage lambda; chemical binding; circular DNA; deoxyribonucleotides; enzyme substrate; ethylene glycols; ionic strengths; molecular biology; oligonucleotides; polymers; virus DNA

A patent covering the polymer-stimulated ligation of DNA has been issued. Macromolecular crowding has been used to obtain efficient phosphorylation by T4 polynucleotide kinase of types of termini in duplex DNA -- such as termini at "nicks" or recessed termini -- which are otherwise only very slowly or incompletely labeled under conventional procedures. Macromolecular crowding was found to extend the range of ionic conditions which support high DNA polymerase reaction rates. High concentrations of nonspecific polymers increased polymerase activity under otherwise inhibitory conditions resulting from relatively high ionic strength. The primary mechanism of the polymer effect seems to be to increase the binding of polymerase to DNA. We have suggested that this effect on protein-DNA complexes is only one example of a general "metabolic buffering" action of crowded solutions on a variety of macromolecular interactions.

一项涉及聚合物刺激 DNA 连接的专利 已发出。 大分子拥挤已被用来获得有效的 T4 多核苷酸激酶对末端类型的磷酸化 双链 DNA——例如“缺口”末端或凹进末端—— 否则只能非常缓慢或不完全地标记 根据常规程序。 发现大分子拥挤可以扩大离子范围 支持高 DNA 聚合酶反应速率的条件。 高浓度的非特异性聚合物增加聚合酶 在其他抑制条件下产生的活性 离子强度相对较高。 其主要机制为 聚合物效应似乎是增加聚合酶的结合 到DNA。 我们认为这种对蛋白质-DNA 的影响 复合物只是一般“代谢缓冲”的一个例子 拥挤溶液对多种大分子的作用 互动。