Y. A. Abramchik, E. A. Zayats, V. I. Timofeev, M. B. Shevtsov, M. A. Kostromina, I. V. Fateev, D. O. Yurovskaya, A. A. Karanov, I. D. Konstantinova, I. P. Kuranova, R. S. Esipov
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引用次数: 0
摘要
摘要 高效生产菌株大肠杆菌 C3030/pET23d+-EcHGPRT，可生产可溶性重组大肠杆菌次黄嘌呤-鸟嘌呤磷酸核糖转移酶（EcHGPRT）。还开发了一种分离和纯化重组蛋白的方法。对天然底物和吡嗪-2-甲酰胺衍生物的特异性活性已经确定。通过毛细管反向扩散法，培育出了适合 X 射线衍射分析的 EcHGPRT 与 3-羟基吡嗪-2-甲酰胺（T-1105）和 6-氟-3-羟基吡嗪-2-甲酰胺（T-705）的复合物晶体。这些晶体属于 P3(1)21；细胞的独立部分包含两个酶分子。
Preliminary X-ray Study of Crystals Obtained by Co-Crystallization of Hypoxanthine‒Guanine Phosphoribosyltransferase from Escherichia coli and Pyrazine-2-Carboxamide Derivatives
A highly effective producer strain Escherichia coli C3030/pET23d+-EcHGPRT, allowing production of recombinant hypoxanthine‒guanine phosphoribosyltransferase from E. coli (EcHGPRT) in a soluble form, has been created. A method for isolating and purifying the recombinant protein has been developed. The specific activity against the natural substrate and pyrazine-2-carboxamide derivatives has been determined. Crystals of the EcHGPRT complexes with 3-hydroxypyrazine-2-carboxamide (T-1105) and 6-fluoro-3-hydroxypyrazine-2-carboxamide (T-705), suitable for X-ray diffraction analysis, have been grown by capillary counter diffusion. X-ray diffraction sets with a resolution of up to 2.4 and 2.5 Å have been collected at the ESRF synchrotron (France, station ID23-1) at a temperature of 100 K. The crystals belong to the sp. gr. P3(1)21; the independent part of the cell contains two enzyme molecules.
期刊介绍：
Crystallography Reports is a journal that publishes original articles short communications, and reviews on various aspects of crystallography: diffraction and scattering of X-rays, electrons, and neutrons, determination of crystal structure of inorganic and organic substances, including proteins and other biological substances; UV-VIS and IR spectroscopy; growth, imperfect structure and physical properties of crystals; thin films, liquid crystals, nanomaterials, partially disordered systems, and the methods of studies.
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