恶性疟原虫DNA聚合酶的纯化与特性分析

Purification and characterization of DNA polymerases from Plasmodium falciparum.

作者信息

Chavalitshewinkoon P, de Vries E, Stam J G, Franssen F F, van der Vliet P C, Overdulve J P

机构信息

Department of Parasitology and Tropical Veterinary Medicine, University of Utrecht, The Netherlands.

出版信息

Mol Biochem Parasitol. 1993 Oct;61(2):243-53. doi: 10.1016/0166-6851(93)90070-e.

DOI:10.1016/0166-6851(93)90070-e

PMID:8264728

Abstract

Fractionation of Plasmodium falciparum cellular extracts by fast protein liquid chromatography (FPLC) identified at least two different DNA polymerases. An aphidicolin-sensitive activity co-purified with a primase activity. This, in combination with other characteristics (processivity, sensitivity to other inhibitors), most likely classifies this enzyme as an alpha-like DNA polymerase. It was, however, relatively resistant to N2-(p-n-butylphenyl)deoxyguanosine 5'-triphosphate (IC50 = 6.6 microM) and differs in this aspect from the host homologue, possibly indicating structural differences between host and parasite DNA polymerase alpha. The other DNA polymerase matched eukaryotic DNA polymerase gamma in all properties tested.

摘要

通过快速蛋白质液相色谱法（FPLC）对恶性疟原虫细胞提取物进行分级分离，鉴定出至少两种不同的DNA聚合酶。一种对阿非迪霉素敏感的活性与一种引发酶活性共纯化。结合其他特性（持续合成能力、对其他抑制剂的敏感性），这种酶很可能被归类为α样DNA聚合酶。然而，它对N2-（对正丁基苯基）脱氧鸟苷5'-三磷酸相对耐药（IC50 = 6.6 microM），在这方面与宿主同源物不同，这可能表明宿主和寄生虫DNA聚合酶α之间存在结构差异。另一种DNA聚合酶在所有测试特性上均与真核生物DNA聚合酶γ匹配。