Crystallization and preliminary X-ray diffraction study of recombinant adenine phosphoribosyltransferase from the thermophilic bacterium Thermus thermophilus strain HB27
- 作者: Sinitsyna E.V.1, Timofeev V.I.1, Tuzova E.S.1, Kostromina M.A.1, Murav’eva T.I.1, Esipov R.S.1, Kuranova I.P.1
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隶属关系:
- Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
- 期: 卷 62, 编号 4 (2017)
- 页面: 580-583
- 栏目: Structure of Macromolecular Compounds
- URL: https://journals.rcsi.science/1063-7745/article/view/191158
- DOI: https://doi.org/10.1134/S106377451704023X
- ID: 191158
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详细
Adenine phosphoribosyltransferase (APRT) belongs to the type I phosphoribosyltransferase family and catalyzes the formation of adenosine monophosphate via transfer of the 5-phosphoribosyl group from phosphoribosyl pyrophosphate to the nitrogen atom N9 of the adenine base. Proteins of this family are involved in a salvage pathway of nucleotide synthesis, thus providing purine base utilization and maintaining the optimal level of purine bases in the body. Adenine phosphoribosyltransferase from the extremely thermophilic Thermus thermophilus strain HB27 was produced using a highly efficient E. coli producer strain and was then purified by affinity and gel-filtration chromatography. This enzyme was successfully employed as a catalyst for the cascade biosynthesis of biologically important nucleotides. The screening of crystallization conditions for recombinant APRT from T. thermophilus HB27 was performed in order to determine the enzyme structure by X-ray diffraction. The crystallization conditions, which were found by the vapor-diffusion technique, were then optimized to apply the counter-diffusion technique. The crystals of the enzyme were grown by the capillary counter-diffusion method. The crystals belong to sp. gr. P1211 and have the following unitcell parameters: a = 69.86 Å, b = 82.16 Å, c = 91.39 Å, α = γ = 90°, β = 102.58°. The X-ray diffraction data set suitable for the determination of the APRT structure at 2.6 Å resolution was collected from the crystals at the SPring-8 synchrotron facility (Japan).
作者简介
E. Sinitsyna
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
编辑信件的主要联系方式.
Email: inna@ns.crys.ras.ru
俄罗斯联邦, Moscow, 117997
V. Timofeev
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: inna@ns.crys.ras.ru
俄罗斯联邦, Moscow, 117997
E. Tuzova
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: inna@ns.crys.ras.ru
俄罗斯联邦, Moscow, 117997
M. Kostromina
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: inna@ns.crys.ras.ru
俄罗斯联邦, Moscow, 117997
T. Murav’eva
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: inna@ns.crys.ras.ru
俄罗斯联邦, Moscow, 117997
R. Esipov
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: inna@ns.crys.ras.ru
俄罗斯联邦, Moscow, 117997
I. Kuranova
Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: inna@ns.crys.ras.ru
俄罗斯联邦, Moscow, 117997
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