INFLUENCES OF MACROMOLECULAR CROWDING ON BIOCHEMICAL SYSTEMS

一项涉及聚合物刺激的DNA连接的专利已经 已经发布了。 大分子拥挤已经被用来获得有效的 T4多核苷酸激酶对细胞内不同类型末端的磷酸化 双链DNA--如“nicks”或凹陷末端-- 否则只能非常缓慢或不完全地标记 在常规程序下。 发现大分子的拥挤扩大了离子的作用范围 支持高DNA聚合酶反应率的条件。 高浓度的非特异性聚合物增加了聚合酶 在其他抑制条件下的活性 离子强度相对较高。其主要作用机制是 聚合物效应似乎增加了聚合酶的结合。 转移到DNA上。我们认为这种对蛋白质-DNA的影响 复合体只是普遍的“代谢缓冲”的一个例子。 拥挤溶液对多种大分子物质的作用 互动。

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.