人肝脏中氨基葡萄糖-6-磷酸N-乙酰基转移酶的纯化、结晶及初步X射线分析
Purification, crystallization and preliminary X-ray analysis of the glucosamine-6-phosphate N-acetyltransferase from human liver.
作者信息
Wang Juan, Zhou Yan-Feng, Li Lan-Fen, Liang Yu-He, Su Xiao-Dong
机构信息
National Laboratory of Protein Engineering and Plant Genetic Engineering, Peking University, Beijing 100871, People's Republic of China.
出版信息
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Nov 1;62(Pt 11):1097-9. doi: 10.1107/S1744309106037730. Epub 2006 Oct 20.
Abstract
Glucosamine-6-phosphate N-acetyltransferase from human liver, which catalyzes the transfer of an acetyl group from acetyl coenzyme A (AcCoA) to the primary amine of D-glucosamine 6-phosphate to form N-acetyl-D-glucosamine 6-phosphate, was expressed in a soluble form from Escherichia coli strain BL21 (DE3). The protein was purified to homogeneity using Ni(2+)-chelating chromatography followed by size-exclusion chromatography. Crystals of the protein were obtained by the hanging-drop vapour-diffusion method and diffracted to 2.6 A resolution. The crystals belonged to space group P4(1)2(1)2 or P4(3)2(1)2, with unit-cell parameters a = b = 50.08, c = 142.88 A.
摘要
人肝脏中的葡糖胺-6-磷酸N-乙酰基转移酶可催化乙酰辅酶A（AcCoA）的乙酰基转移至D-葡糖胺6-磷酸的伯胺上，形成N-乙酰-D-葡糖胺6-磷酸，该酶以可溶形式在大肠杆菌BL21（DE3）菌株中表达。通过镍（2+）螯合层析，随后进行尺寸排阻层析，将该蛋白纯化至同质。通过悬滴气相扩散法获得了该蛋白的晶体，其衍射分辨率达到2.6 Å。晶体属于空间群P4(1)2(1)2或P4(3)2(1)2，晶胞参数a = b = 50.08，c = 142.88 Å。
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