嗜热栖热菌HB8 MutM蛋白参与氧化DNA损伤修复的结晶及初步X射线晶体学研究。

Crystallization and preliminary X-ray crystallographic studies of Thermus thermophilus HB8 MutM protein involved in repairs of oxidative DNA damage.

作者信息

Sugahara M, Mikawa T, Kato R, Fukuyama K, Kumasaka T, Yamamoto M, Inoue Y, Kuramitsu S

机构信息

Department of Biology, Graduate School of Science, Osaka University, Toyonaka, Osaka 560-0043, Japan.

出版信息

J Biochem. 2000 Jan;127(1):9-11. doi: 10.1093/oxfordjournals.jbchem.a022588.

DOI:10.1093/oxfordjournals.jbchem.a022588

PMID:10731660

Abstract

MutM protein, which removes the oxidatively damaged DNA base product, 8-oxoguanine (GO), has been crystallized by means of a hanging-drop vapor-diffusion procedure using polyethyleneglycol monomethylether 2000 as a precipitant in 2-(cyclohexylamino) ethanesulfonic acid (CHES) buffer, pH 9.8. The diffraction data derived from oscillation photographs indicate that the crystals belong to the monoclinic system and space group P2(1). The crystals have unit-cell dimensions of a = 45.4 A, b = 62.0 A, c = 99.7 A, and beta = 90.8 degrees. Assuming that the asymmetric unit contains two molecules, the Vm value was calculated to be 2.35 A(3).Da(-1). The crystals diffracted X-rays to at least 2.1 A resolution and were suitable for high-resolution X-ray crystal structure determination.

摘要

MutM蛋白可去除经氧化损伤的DNA碱基产物8-氧代鸟嘌呤（GO），该蛋白已通过悬滴气相扩散法进行结晶，使用聚乙二醇单甲醚2000作为沉淀剂，在pH值为9.8的2-（环己基氨基）乙烷磺酸（CHES）缓冲液中进行。从振荡照片获得的衍射数据表明，晶体属于单斜晶系，空间群为P2(1)。晶体的晶胞参数为a = 45.4 Å，b = 62.0 Å，c = 99.7 Å，β = 90.8°。假设不对称单位包含两个分子，计算得到的Vm值为2.35 Å(3).Da(-1)。这些晶体对X射线的衍射分辨率至少为2.1 Å，适合进行高分辨率X射线晶体结构测定。