Preliminary X-ray Study of Crystals Obtained by Co-Crystallization of Hypoxanthine‒Guanine Phosphoribosyltransferase from Escherichia coli and Pyrazine-2-Carboxamide Derivatives

IF 0.6 4区材料科学 Q4 CRYSTALLOGRAPHY Crystallography Reports Pub Date : 2024-02-05 DOI:10.1134/s1063774523600965

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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Abstract

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.

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大肠杆菌次黄嘌呤-鸟嘌呤磷酸核糖基转移酶与吡嗪-2-甲酰胺衍生物共结晶所获晶体的初步 X 射线研究

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。

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来源期刊

Crystallography Reports 化学-晶体学

CiteScore

1.10

自引率

28.60%

发文量

审稿时长

4-8 weeks

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.