Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human β-galactosidase
Expression, purification, crystallization and preliminary X-ray crystallographic analysis of human β-galactosidase
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人β-半乳糖苷酶的表达、纯化、结晶及初步X射线晶体分析
DOI：
10.1107/s1744309111047920
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发表时间：
2012
期刊：
影响因子：
--
通讯作者：
Y.
中科院分区：
文献类型：
--
作者：
Usui;K.;Ohto;U.;Ochi;T.;Shimizu;T.;Satow;Y.
β-d-半乳糖苷酶（β-Gal）是一种外切糖苷酶，其从糖蛋白、鞘脂和硫酸角质素切割β-半乳糖苷。本文报道了人溶酶体β-Gal的表达、纯化、结晶和初步的X-射线晶体学分析。采用坐滴气相扩散法结晶β-Gal与其产物半乳糖和抑制剂1-脱氧半乳糖野尻霉素的复合物。所得晶体为类质同晶，属于空间群P21。β-Gal-半乳糖和β-Gal-抑制剂复合物的晶胞参数分别为a = 94.8，B = 116.1，c = 140.3 nm，β = 92.2°和a = 94.8，B = 116.0，c = 140.3 nm，β = 92.2°。 收集两种晶体的衍射数据至1.8 μ m分辨率。
β-d-Galactosidase (β-Gal) is an exoglycosidase that cleaves β-galactosides from glycoproteins, sphingolipids and keratan sulfate. This study reports the expression, purification, crystallization and preliminary X-ray crystallographic analysis of human lysosomal β-Gal. The sitting-drop vapour-diffusion method was used to crystallize β-Gal in complexes with its product galactose and with the inhibitor 1-deoxygalactonojirimycin. The resulting crystals were isomorphous and belonged to space group P21. The crystals of the β-Gal–galactose and the β-Gal–inhibitor complexes had unit-cell parameters a = 94.8, b = 116.1, c = 140.3 Å, β = 92.2° and a = 94.8, b = 116.0, c = 140.3 Å, β = 92.2°, respectively. Diffraction data were collected to 1.8 Å resolution for both crystals.