Preliminary X-ray Study of Crystals Obtained by Co-Crystallization of Hypoxanthine‒Guanine Phosphoribosyltransferase from Escherichia coli and Pyrazine-2-Carboxamide Derivatives-Reference-Cited by-同舟云学术

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

Published:2023-11-01 Issue:6 Volume:68 Page:854-858
ISSN:0023-4761
Container-title:Кристаллография
language:
Short-container-title:Kristallografiâ

Author:

Abramchik Yu. A.¹²,Zayats E. A.¹,Timofeev V. I.²³,Shevtsov M. B.⁴,Kostromina M. A.¹,Fateev I. V.¹,Yurovskaya D. O.¹,Karanov A. A.¹,Konstantinova I. D.¹,Kuranova I. P.²³,Esipov R. S.¹

Affiliation:

1. Shemyakin‒Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997, Moscow, Russia

2. Shubnikov Institute of Crystallography, Federal Research Centre “Crystallography and Photonics,” Russian Academy of Sciences, 119333, Moscow, Russia

3. National Research Centre “Kurchatov Institute”, 123182, Moscow, Russia

4. Moscow Institute of Physics and Technology, 141700, Dolgoprudnyi, Moscow oblast, Russia

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.

Publisher

The Russian Academy of Sciences

Reference26 articles.

1. Hucke F.I.L., Bestehorn-Willmann M., Bassetto M. et al. // Virus Genes. 2022. V. 58 (3). P. 188. https://doi.org/10.1007/s11262-022-01892-x

2. De Vleeschauwer A.R., Lefebvre D.J., Willems T. et al. // Transbound. Emerg. Dis. 2016. V. 63 (2). P. e205. https://doi.org/10.1111/tbed.12255

3. Hayden F.G., Shindo N. // Curr. Opin. Infect. Dis. 2019. V. 32 (2). P. 176. https://doi.org/10.1097/QCO.0000000000000532

4. Delang L., Abdelnabi R., Neyts J. // Antiviral Res. 2018. V. 153. P. 85. https://doi.org/10.1016/j.antiviral.2018.03.003

5. De Clercq E., Li G. // Clin. Microbiol. Rev. 2016. V. 29 (3). P. 695. https://doi.org/10.1128/CMR.00102-15