通过色谱法和电泳法对真核生物DNA拓扑异构酶进行分离和功能分析。

Separation and functional analysis of eukaryotic DNA topoisomerases by chromatography and electrophoresis.

作者信息

Knudsen B R, Straub T, Boege F

机构信息

Department of Molecular and Structural Biology, Univ. Aarhus, Denmark.

出版信息

J Chromatogr B Biomed Appl. 1996 Sep 20;684(1-2):307-21. doi: 10.1016/0378-4347(96)00152-1.

DOI:10.1016/0378-4347(96)00152-1

PMID:8906479

Abstract

DNA topoisomerases are enzymes that control DNA topology by cleaving and rejoining DNA strands and passing other DNA strands through the transient gaps. Consequently, these enzymes play a crucial role in the regulation of the physiological function of the genome. Beyond their normal functions, topoisomerases are important cellular targets in the treatment of human cancers. In this review we summarize current protocols for extracting and purifying DNA topoisomerases, and for separating subtypes and isoforms of these enzymes. Furthermore, we discuss methods for measuring the catalytic activity of topoisomerases and for monitoring the molecular effects of topoisomerase-directed antitumor drugs in cell-free assays.

摘要

DNA拓扑异构酶是一类通过切割和重新连接DNA链并使其他DNA链穿过瞬时缺口来控制DNA拓扑结构的酶。因此，这些酶在基因组生理功能的调节中起着关键作用。除了其正常功能外，拓扑异构酶是人类癌症治疗中的重要细胞靶点。在本综述中，我们总结了目前提取和纯化DNA拓扑异构酶、分离这些酶的亚型和同工型的方法。此外，我们还讨论了在无细胞分析中测量拓扑异构酶催化活性以及监测拓扑异构酶导向抗肿瘤药物分子效应的方法。