Bulletin of the Korean Chemical Society

Korean Chemical Society (대한화학회)
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Mutagenic Analysis of hPNMT Confirms the Importance of Lys57 and the Inhibitor Binding Site

  • Jeong, Ki-Woong (Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University) ;
  • Kang, Dong-Il (Department of Chemistry, Konkuk University) ;
  • Lee, Jee-Young (Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University) ;
  • Kim, Yang-Mee (Department of Bioscience and Biotechnology, Bio/Molecular Informatics Center, Konkuk University)
  • Received : 2010.11.10
  • Accepted : 2010.11.27
  • Published : 2011.02.20
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Abstract

In previous report, with the aid of receptor-oriented pharmacophore-based in silico screening, we characterized three novel hPNMT inhibitors (YPN010, YPN016, and YPN017) and proposed that the hydrogen bonding interaction between inhibitors and side chain of Lys57 is very important to inhibitory activity of hPNMT. To confirm the importance of Lys57, mutant with substitution of Lys57 with Ala was cloned and binding study was performed for a K57A mutant of hPNMT using STD-NMR and fluorescence experiments. The binding constants for three novel inhibitors with mutant hPNMT were dramatically decreased compared to those with wild-type protein. K57A mutant-induced conversion of noradrenaline to adrenaline was suppressed about 95 % compared to wild-type hPNMT. Mutagenic analysis using a K57A mutant confirmed the importance of the Lys57 residue in binding of the inhibitor candidate to hPNMT as well as enzymatic activity of hPNMT, implying that these results are consistent with our binding model.

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References

  1. Matsubara, K.; Aoyama, K.; Suno, M.; Awaya, T. Neurotoxicol. Teratol. 2002, 24, 593. https://doi.org/10.1016/S0892-0362(02)00212-X
  2. Berecek, K. H.; Brody, M. J. Am. J. Physiol. 1982, 242, H593.
  3. Martin, J. L.; Begun, J.; McLeish, M. J.; Caine, J. M.; Grunewald,G. L. Structure 2001, 9, 977. https://doi.org/10.1016/S0969-2126(01)00662-1
  4. Axelrod, J. J. Biol. Chem. 1962, 237, 1657.
  5. Wu, Q.; Gee, C. L.; Lin, F.; Tyndall, J. D.; Martin, J. L.; Grunewald,G. L.; McLeish, M. J. J. Med. Chem. 2005, 48, 7243. https://doi.org/10.1021/jm050568o
  6. Grunewald, G. L.; Seim, M. R.; Bhat, S. R.; Wilson, M. E.; Criscione,K. R. Bioorg. Med. Chem. 2008, 16, 542. https://doi.org/10.1016/j.bmc.2007.08.066
  7. Grunewald, G. L.; Seim, M. R.; Regier, R. C.; Criscione, K. R.Bioorg. Med. Chem. 2008, 15, 1298.
  8. Grunewald, G. L.; Seim, M. R.; Regier, R. C.; Martin, J. L.; Gee,C. L.; Drinkwater, N.; Criscione, K. R. J. Med. Chem. 2006, 49,5424. https://doi.org/10.1021/jm060466d
  9. Grunewald, G. L.; Lu, J.; Criscione, K. R.; Okoro, C. O. Bioorg. Med. Chem. Lett. 2005, 15, 5319. https://doi.org/10.1016/j.bmcl.2005.08.033
  10. Grunewald, G. L.; Romero, F. A.; Criscione, K. R. J. Med. Chem.2005, 48, 1806. https://doi.org/10.1021/jm049594x
  11. Kang, D. I.; Lee, J. Y.; Kim, W.; Jeong, K. W.; Shin, S.; Yang, J.;Park, E.; Chae, Y. K.; Kim, Y. Mol. Cells 2010, 29, 595. https://doi.org/10.1007/s10059-010-0074-3
  12. Nissler, L.; Gebhardt, R.; Berger, S. Anal. Bioanal. Chem. 2004,379, 1045.
  13. Mishra, B.; Barik, A.; Priyadarsini, K. I.; Mohan, H. J. Chem. Sci.2005, 117, 641. https://doi.org/10.1007/BF02708293
  14. Azani, M-R.; Hassanpour, A.; Bordbar, A-K.; Mirkhani, V. Bull. Korean Chem. Soc. 2009, 30, 1973. https://doi.org/10.5012/bkcs.2009.30.9.1973
  15. Mayer, M.; Meyer, B. J. Am. Chem. Soc. 2001, 123, 6108. https://doi.org/10.1021/ja0100120
  16. Macnaughtan, M. A.; Kamar, M.; Alvarez-Manilla, G.; Venot,A.; Glushka, J.; Pierce, J. M.; Prestegard, J. H. J. Mol. Biol. 2006,366, 1266.
  17. Gee, C. L.; Tyndall, J. D.; Grunewald, G. L.; Wu, Q.; McLeish,M. J.; Martin, J. L. Biochemistry 2005, 44, 16875. https://doi.org/10.1021/bi051636b
  18. Krovat, E. M.; Frühwirth, K. H.; Langer, T. J. Chem. Inf. Model.2005, 45, 146. https://doi.org/10.1021/ci049778k
  19. Gee, C. L.; Drinkwater, N.; Tyndall, J. D.; Grunewald, G. L.; Wu,Q.; McLeish, M. J.; Martin, J. L. J. Med. Chem. 2007, 50, 4845. https://doi.org/10.1021/jm0703385
  20. Lee, J. Y.; Jeong, K. W.; Kim, Y. Bull. Korean Chem. Soc. 2009,30, 1835. https://doi.org/10.5012/bkcs.2009.30.8.1835
  21. Grunewald, G. L.; Seim, M. R.; Regier, R. C.; Martin, J. L.; Gee,C. L.; Drinkwater, N.; Criscione, K. R. J. Med. Chem. 2006, 49,5424. https://doi.org/10.1021/jm060466d
  22. Grunewald, G. L.; Borchardt, R. T.; Rafferty, M. F.; Krass, P. Mol. Pharmacol. 1981, 20, 377.