刚地弓形虫尿嘧啶磷酸核糖转移酶的表达、纯化、结晶及初步X射线衍射分析
Expression, purification, crystallization and preliminary X-ray diffraction analysis of uracil phosphoribosyltransferase of Toxoplasma gondii.
作者信息
Barchue J, Symersky J, Narayana S V, Moore J K, DeLucas L J, Chattopadhyay D
机构信息
Center for Macromolecular Crystallography, University of Alabama at Birmingham, 1918 University Boulevard, Birmingham, AL 35294, USA.
出版信息
Acta Crystallogr D Biol Crystallogr. 1999 Jan;55(Pt 1):347-9. doi: 10.1107/S090744499800821X. Epub 1999 Jan 1.
Recombinant uracil phosphoribosyltransferase (UPRT) enzyme of Toxoplasma gondii was expressed in Escherichia coli and purified from the cell-free extract by a combination of chromatographic steps. The recombinant protein was enzymatically active when tested in an in vitro UPRT assay. The purified protein was crystallized using the hanging-drop vapor-diffusion technique with ammonium phosphate as precipitant. The crystallized protein also exhibited UPRT activity. Crystals diffract to 2.4 A resolution and belong to space group P3121 or P3221 with unit-cell dimensions a = b = 119.9, c = 70.8 A and two molecules per asymmetric unit.
弓形虫重组尿嘧啶磷酸核糖转移酶（UPRT）在大肠杆菌中表达，并通过一系列色谱步骤从无细胞提取物中纯化出来。在体外UPRT测定中测试时，重组蛋白具有酶活性。使用以磷酸铵为沉淀剂的悬滴气相扩散技术对纯化后的蛋白进行结晶。结晶后的蛋白也表现出UPRT活性。晶体衍射分辨率为2.4 Å，属于空间群P3121或P3221，晶胞参数a = b = 119.9，c = 70.8 Å，每个不对称单元中有两个分子。
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