Abstract
Pyrimidine nucleoside phosphorylases, which are widely used in the biotechnological production of nucleosides, have different substrate specificity for pyrimidine nucleosides. An interesting feature of these enzymes is that the three-dimensional structure of thymidine-specific nucleoside phosphorylase is similar to the structure of nonspecific pyrimidine nucleoside phosphorylase. The three-dimensional structures of thymidine phosphorylase from Salmonella typhimurium and nonspecific pyrimidine nucleoside phosphorylase from Bacillus subtilis in complexes with a sulfate anion were determined for the first time by X-ray crystallography. An analysis of the structural differences between these enzymes demonstrated that Lys108, which is involved in the phosphate binding in pyrimidine nucleoside phosphorylase, corresponds to Met111 in thymidine phosphorylases. This difference results in a decrease in the charge on one of the hydroxyl oxygens of the phosphate anion in thymidine phosphorylase and facilitates the catalysis through SN2 nucleophilic substitution. Based on the results of X-ray crystallography, the virtual screening was performed for identifying a potent inhibitor (anticancer agent) of nonspecific pyrimidine nucleoside phosphorylase, which does not bind to thymidine phosphorylase. The molecular dynamics simulation revealed the stable binding of the discovered compound—2-pyrimidin-2-yl-1H-imidazole-4-carboxylic acid—to the active site of pyrimidine nucleoside phosphorylase.
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References
T. Utagawa, J. Mol. Catal. B 6 (3), 215 (1999).
M. J. Pugmire and S. E. Ealick, Biochem J. 361 (1), 1 (2002).
J. Vande Voorde, S. Liekens, F. Gago, et al., Nucleosides, Nucleotides Nucleic Acids 33 (4–6), 394 (2014).
T. Hamamoto, T. Noguchi, and Y. Midorikawa, Biosci. Biotechnol. Biochem. 60 (7), 1179 (1996).
N. G. Panova, C. S. Alexeev, A. S. Kuzmichov, et al., Biochemistry (Moscow) 72 (1), 21 (2007).
J. Vande Voorde, F. Gago, K. Vrancken, et al., Biochem. J. 445 (1), 113 (2012).
M. J. Pugmire, W. J. Cook, A. Jasanoff, et al., J. Mol. Biol. 281 (2), 285 (1998).
M. J. Pugmire and S. E. Ealick, Structure 6 (11), 1467 (1998).
S. W. Rick, Y. G. Abashkin, R. L. Hilderbrandt, et al., Proteins 37 (2), 242 (1999).
V. V. Balaev, A. A. Lashkov, A. G. Gabdulkhakov, et al., Acta Crystallogr. F 72 (3), 224 (2016).
V. Timofeev, Y. Abramchik, N. Zhukhlistova, et al., Acta Crystallogr. D 70 (4), 1155 (2014).
R. A. Norman, S. T. Barry, M. Bate, et al., Structure 12 (1), 75 (2004).
K. El Omari, A. Bronckaers, S. Liekens, et al., Biochem. J. 399 (2), 199 (2006).
A. Bronckaers, F. Gago, J. Balzarini, et al., Med. Res. Rev. 29 (6), 903 (2009).
W. Kabsch, Acta Crystallogr. D 66 (2), 125 (2010).
A. J. McCoy, Acta Crystallogr. D 63 (1), 32 (2007).
P. D. Adams, P. V. Afonine, G. Bunkoczi, et al., Acta Crystallogr. D 66 (2), 213 (2010).
P. V. Afonine, R. W. Grosse-Kunstleve, N. Echols, et al., Acta Crystallogr. D 68 (4), 352 (2012).
G. N. Murshudov, A. A. Vagin, and E. J. Dodson, Acta Crystallogr. D 53 (3), 240 (1997).
P. Emsley and K. Cowtan, Acta Crystallogr. D 60 (12), 2126 (2004).
P. Emsley, B. Lohkamp, W. G. Scott, et al., Acta Crystallogr. D 66 (4), 486 (2010).
I. W. Davis, A. Leaver-Fay, V. B. Chen, et al., Nucleic Acids Res. 35, Web Server Issue, 375 (2007).
D. Van Der Spoel, E. Lindahl, B. Hess, et al., J. Comput. Chem. 26 (16), 1701 (2005).
T. Schlesier and G. Diezemann, J. Phys. Chem. B 117 (6), 1862 (2013).
M. R. Walter, W. J. Cook, L. B. Cole, et al., J. Biol. Chem. 265 (23), 14016 (1990).
F. Sievers, A. Wilm, D. Dineen, et al., Mol. Syst. Biol. 7, 539 (2011).
W. G. Touw, C. Baakman, J. Black, et al., Nucleic Acids Res. 43, Database Issue, 364 (2015).
E. Mitsiki, A. C. Papageorgiou, S. Iyer, et al., Biochem. Biophys. Res. Commun. 386 (4), 666 (2009).
M. H. Iltzsch, M. H. el Kouni, and S. Cha, Biochemistry 24 (24), 6799 (1985).
S. Liekens, A. I. Hernandez, D. Ribatti, et al., J. Biol. Chem. 279 (28), 29598 (2004).
D. W. Cruickshank, Acta Crystallogr. D 55 (3), 583 (1999).
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Original Russian Text © V.V. Balaev, A.A. Lashkov, I.I. Prokofev, A.G. Gabdulkhakov, T.A. Seregina, A.S. Mironov, C. Betzel, A.M. Mikhailov, 2016, published in Kristallografiya, 2016, Vol. 61, No. 5, pp. 797–808.
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Balaev, V.V., Lashkov, A.A., Prokofev, I.I. et al. Substrate specificity of pyrimidine nucleoside phosphorylases of NP-II family probed by X-ray crystallography and molecular modeling. Crystallogr. Rep. 61, 830–841 (2016). https://doi.org/10.1134/S1063774516050023
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DOI: https://doi.org/10.1134/S1063774516050023