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Three-Dimensional Structure of Recombinant Adenine Phosphoribosyltransferase from Thermophilic Bacterial Strain Thermus thermophilus HB27

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Abstract

Adenine phosphoribosyltransferase (APRT) from a thermophilic bacterial strain Thermus thermophilus НВ27 (TthHB27APRT) belongs to the family of type I phosphoribosyltransferases and catalyzes the magnesium-dependent transfer of 5'-phosphoribosyl group from 5'-phosphoribosylpyrophosphate to N9 adenine nitrogen with formation of adenosine-5'-monophosphate and pyrophosphate. The crystals of the recombinant enzyme suitable for X-ray study were grown in a capillary using the counter-diffusion technique. Crystals with unit-cell parameters α = 69.860 Å, b = 82.160 Å, c = 91.390 Å, α = 90.00°, β = 102.58°, and γ = 90.00° belong to the space group Р21 and contain six enzyme monomers in the asymmetric unit. The set of X-ray data from grown crystals was collected on a Spring-8 synchrotron radiation facility (Japan) and three-dimensional structure of the enzyme was solved at 2.7-Å resolution by molecular replacement method using the BALBES software. The polypeptide fold in the enzyme monomer and the structure of biologically active dimer were described. Based on the comparison with structures of homologous APRTs from a thermophilic strain ThtHB8 and Homo sapiens, positions of active site and a number of functionally important amino acids were located.

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Abbreviations

PRT:

phosphoribosyltransferase

APRT:

adenine phosphoribosyltransferase

TthHB27APRt and Tth-HB8APRT:

adenine phosphoribosyltransferases of the thermophilic bacteria Thermus thermophilus strains HB27 and HB8

HsAPRT:

adenine phosphoribosyltransferase from Homo sapiens

PRPP:

5'-phosphoribosyl-1-pyrophosphate

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Authors and Affiliations

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

    R. S. Esipov, V. I. Timofeev, E. V. Sinitsyna, E. S. Tuzova, L. V. Esipova, M. A. Kostromina, I. P. Kuranova & A. I. Miroshnikov

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  1. R. S. Esipov
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  2. V. I. Timofeev
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  3. E. V. Sinitsyna
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  4. E. S. Tuzova
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  5. L. V. Esipova
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  6. M. A. Kostromina
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  7. I. P. Kuranova
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  8. A. I. Miroshnikov
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Correspondence to R. S. Esipov.

Additional information

Original Russian Text © R.S. Esipov, V.I. Timofeev, E.V. Sinitsyna, E.S. Tuzova, L.V. Esipova, M.A. Kostromina, I.P. Kuranova, A.I. Miroshnikov, 2018, published in Bioorganicheskaya Khimiya, 2018, Vol. 44, No. 5, pp. 506–513.

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Esipov, R.S., Timofeev, V.I., Sinitsyna, E.V. et al. Three-Dimensional Structure of Recombinant Adenine Phosphoribosyltransferase from Thermophilic Bacterial Strain Thermus thermophilus HB27. Russ J Bioorg Chem 44, 504–510 (2018). https://doi.org/10.1134/S1068162018050047

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  • DOI: https://doi.org/10.1134/S1068162018050047

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