Substrate Specificity of Protein Kinase UL97, an antiviral target, on Mutant Peptide Substrates Derived from a Peptide, KESYSVYVYKV

KESYSVYVYKV로부터 변형된 펩타이드 기질을 이용한 항바이러스제의 타깃이 되는 UL97 단백질 인산화 효소의 기질 특이성

  • Baek, Moon-Chang (Department of Molecular Medicine, School of Medicine, Kyungpook National University)
  • 백문창 (경북대학교 의과대학 분자의학교실)
  • Published : 2008.12.31

Abstract

Human cytomegalovirus expresses an unusual protein kinase UL97, a member of ${H_V}{U_L}$ family of protein kinase. UL97 can phosphorylate nucleoside analogs such as ganciclovir as well as protein/peptide. It has previously been reported that UL97 is able to phosphorylate a KESYSVYVYKV peptide and that P+5 position (K) is important. We examined the extent of contribution of other positions (P-4 through P+6) of the peptide to be substrate of UL97 using alanine substituted peptides (Ala scanning) and deleted peptides. The result suggested that the E (P-2) is negative effect and P+5 (K) is still important. The peptide YSVYVYK is the shortest substrate enough to show high activity, which could be a starting point to develop peptidomimetic drug. This study would give important information to deeply understand the substrate specificity of UL97 and develop an antiviral drug using the small peptide identified here.

Keywords

References

  1. Littler, E., Stuart, A. D. and Chee, M. S. : Human cytomegalovirus UL97 open reading frame encodes a protein that phosphorylates the antiviral nucleoside analogue ganciclovir. Nature 358, 160 (1992) https://doi.org/10.1038/358160a0
  2. Sullivan, V., Talarico, C. L., Stanat, S. C., Davis, M., Coen, D. M. and Biron, K. K. : A protein kinase homologue controls phosphorylation of ganciclovir in human cytomegalovirus-infected cells. Nature 358, 162 (1992) https://doi.org/10.1038/358162a0
  3. Krosky, P. M., Baek, M. C. and Coen, D. M. : The human cytomegalovirus UL97 protein kinase, an antiviral drug target, is required at the stage of nuclear egress. J. Virol. 77, 905 (2003) https://doi.org/10.1128/JVI.77.2.905-914.2003
  4. Wolf, D. G., Courcelle, C. T., Prichard, M. N. and Mocarski, E. S. : Distinct and separate roles for herpesvirus-conserved UL97 kinase in cytomegalovirus DNA synthesis and encapsidation. Proc. Natl. Acad. Sci. U.S.A. 98, 1895 (2001)
  5. Biron, K. K., Harvery, R. J., Chamberlain, S. C., Good, S. S., Smith, A. A. 3rd, davis, M. G., Talarico, C. L., Miller, W. H., Ferris, R., Dornsife, R. E., Stanat, S. C., Drach, J. C., Townsed, L. B. and Koszalka, G. W. : Potent and selective inhibition of human cytomegalovirus replication by 1263W94, a benzimidazole L-riboside with a unique mode of action. Antimicrob. Agents Chemother. 46, 2365 (2002) https://doi.org/10.1128/AAC.46.8.2365-2372.2002
  6. Talarico, C. L., Burnette, T. C., Miller, W. H., Smith, S. L., Davis, M. G., Stanat, S. C., Ng, T. I., He, Z., Coen, D. M., Roizman, B. and Biron, K. K. : Acyclovir is phosphorylated by the human cytomegalovirus UL97 protein. Antimicrob. Agents Chemother. 43, 1941 (1999)
  7. Baek, M. C., Krosky, P. M., He, Z. and Coen, D. M. : Specific phosphorylation of exogenous protein and peptide substrates by the human cytomegalovirus UL97 protein kinase. Importance of the P+5 position. J. Biol. Chem. 277, 29593 (2002) https://doi.org/10.1074/jbc.M202312200
  8. Krosky, P. M., Baek, M. C., Jahng, W. J., Barrera, I., Harvey, R. J., Biron, K. K., Coen, D. M. and Sethna, P. B. : The human cytomegalovirus UL44 protein is a substrate for the UL97 protein kinase. J. Virol. 77, 7720 (2003) https://doi.org/10.1128/JVI.77.14.7720-7727.2003
  9. Baek, M. C., Krosky, P. M. and Coen, D. M. : Relationship between autophosphorylation and phosphorylation of exogenous substrates by the human cytomegalovirus UL97 protein kinase. J. Virol. 76, 11943 (2002) https://doi.org/10.1128/JVI.76.23.11943-11952.2002
  10. Prichard, M. N., Gao, N., Jairath, S., Mulamba, G., Krosky, P., Coen, D. M., Parker, B. O. and Pari, G. S. : A recombinant human cytomegalovirus with a large deletion in UL97 has a severe replication deficiency. J. Virol. 73, 5663 (1999)
  11. Chee, M. S., Lawrence, G. L. and Barrell, B. G. : Alpha-, beta- and gammaherpesviruses encode a putative phosphotransferase. J. Gen. Virol. 70, 1151 (1989) https://doi.org/10.1099/0022-1317-70-5-1151
  12. Hanks, S. K. and Hunter, T. : Protein kinases 6. The eukaryotic protein kinase superfamily: kinase (catalytic) domain structure and classification. FASEB J. 9, 576 (1995) https://doi.org/10.1096/fasebj.9.8.7768349
  13. Crossen, R. and Gruenwald, S. : In Baculovirus Expression Vector System Manual, 5th ed. Pharmingen, San Diego (1998)
  14. He, Z., Kim, Y., Chu, L., Ohmstede, C., Biron, K. K. and Coen, D. M. : The human cytomegalovirus UL97 protein is a protein kinase that autophosphorylates on serines and threonines. J. Virol. 71, 405 (1997)