嗜热栖热放线菌7株尿嘧啶-DNA糖基化酶的纯化、结晶及初步X射线分析
Purification, crystallization and preliminary X-ray analysis of uracil-DNA glycosylase from Sulfolobus tokodaii strain 7.
作者信息
Kawai Akito, Higuchi Shigesada, Tsunoda Masaru, Nakamura Kazuo T, Miyamoto Shuichi
机构信息
Faculty of Pharmaceutical Sciences, Sojo University, 4-22-1 Ikeda, Kumamoto 860-0082, Japan.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Sep 1;68(Pt 9):1102-5. doi: 10.1107/S1744309112030278. Epub 2012 Aug 31.
Abstract
Uracil-DNA glycosylase (UDG) specifically removes uracil from DNA by catalyzing hydrolysis of the N-glycosidic bond, thereby initiating the base-excision repair pathway. Although a number of UDG structures have been determined, the structure of archaeal UDG remains unknown. In this study, a deletion mutant of UDG isolated from Sulfolobus tokodaii strain 7 (stoUDGΔ) and stoUDGΔ complexed with uracil were crystallized and analyzed by X-ray crystallography. The crystals were found to belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 52.2, b = 52.3, c = 74.7 Å and a = 52.1, b = 52.2, c = 74.1 Å for apo stoUDGΔ and stoUDGΔ complexed with uracil, respectively.
摘要
尿嘧啶-DNA糖基化酶（UDG）通过催化N-糖苷键的水解从DNA中特异性去除尿嘧啶，从而启动碱基切除修复途径。尽管已经确定了许多UDG的结构，但古细菌UDG的结构仍然未知。在本研究中，对从嗜热栖热放线菌7株（stoUDGΔ）分离的UDG缺失突变体以及与尿嘧啶复合的stoUDGΔ进行了结晶，并通过X射线晶体学进行分析。发现晶体属于正交空间群P2(1)2(1)2(1)，apo stoUDGΔ和与尿嘧啶复合的stoUDGΔ的晶胞参数分别为a = 52.2，b = 52.3，c = 74.7 Å和a = 52.1，b = 52.2，c = 74.1 Å。
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