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克氏锥虫核苷二磷酸激酶1的蛋白质制备、结晶及初步X射线分析

Protein preparation, crystallization and preliminary X-ray analysis of Trypanosoma cruzi nucleoside diphosphate kinase 1.

作者信息

Gómez Barroso J A, Pereira H, Miranda M, Pereira C, Garratt R C, Aguilar C F

机构信息

Laboratorio de Biología Molecular Estructural, Instituto Multidisciplinarlo de Investigación en Biología, Universidad Nacional de San Luis and CONICET, Avenida Ejército de los Andes 950, Bloque 1, San Luis, Argentina.

出版信息

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Jul 1;66(Pt 7):862-5. doi: 10.1107/S1744309110013886. Epub 2010 Jun 26.

Abstract

The flagellated protozoan parasite Trypanosoma cruzi is the aetiological agent of Chagas disease. Nucleoside diphosphate kinases (NDPKs) are enzymes that are involved in energy management and nucleoside balance in the cell. T. cruzi TcNDPK1, a canonical isoform, was overexpressed in Escherichia coli as an N-terminally poly-His-tagged fusion protein and crystallized. Crystals grew after 72 h in 0.2 M MgCl(2), 20% PEG 3350. Data were collected to 3.5 A resolution using synchrotron X-ray radiation at the National Synchrotron Light Laboratory (Campinas, Brazil). The crystals belonged to the trigonal space group P3, with unit-cell parameters a = b = 127.84, c = 275.49 A. Structure determination is under way and will provide relevant information that may lead to the first step in rational drug design for the treatment of Chagas disease.

摘要

有鞭毛的原生动物寄生虫克氏锥虫是恰加斯病的病原体。核苷二磷酸激酶(NDPKs)是参与细胞能量管理和核苷平衡的酶。克氏锥虫的TcNDPK1是一种典型的同工型,作为N端带有多聚组氨酸标签的融合蛋白在大肠杆菌中过表达并结晶。晶体在0.2 M MgCl₂、20% PEG 3350中培养72小时后生长。使用巴西坎皮纳斯国家同步辐射光实验室的同步辐射X射线辐射收集了分辨率为3.5 Å的数据。晶体属于三方晶系空间群P3,晶胞参数a = b = 127.84,c = 275.49 Å。结构测定正在进行中,将提供可能导致恰加斯病合理药物设计第一步的相关信息。

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Parasitol Today. 1998 Jul;14(7):265-72. doi: 10.1016/s0169-4758(98)01263-0.
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