약학회지 (YAKHAK HOEJI)

  • 제50권4호
  • /
  • Pages.268-271
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  • 2006
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  • 0377-9556(pISSN)
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  • 2383-9457(eISSN)
대한약학회 (The Pharmaceutical Society of Korea)
권호 표지

Spot Assay를 통한 Human Cytomegalovirus의 UL97 단백질 인산화 효소의 기질 특이성

Substrate Specificity of UL97 Protein Kinase from Human Cytomegalovirus using Spot Assay

  • 백문창 (경북대학교 의과대학 분자의학교실)
  • Baek, Moon-Chang (Department of Molecular Medicine, School of Medicine, Kyungpook National University)
  • 발행 : 2006.08.31
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초록

Protein kinase UL97 is an unusual protein kinase that can phosphorylate nucleoside analogs as well as protein/peptide. Previously we found a H2B-derived peptide, KESYSVYVYKV and reported that the P+5 position (K) is important. To further understand the substrate specificity at the P+5 position, we introduced spot assay system and showed that a peptide containing K residue among other amino acids at the P+5 position is the best substrate. Also other residues such as M, I, L, or G are good enough to be substrate of UL97. This result may aid the discovery of a new antiviral inhibitor.

키워드

참고문헌

  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 cytomegalovirusinfected cells. Nature 358, 162 (1992) https://doi.org/10.1038/358162a0
  3. 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)
  4. 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
  5. 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)
  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. 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
  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., 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
  10. Selleseth, D. W., Talarico, C. L., Miller, T., Lutz, M. W., Biron, K. K. and Harvey, R. J. : In vitro activities of benzimidazole D- and L-ribonucleosides against herpesviruses. Antimicrob. Agents Chemother. 47, 2186 (2003) https://doi.org/10.1128/AAC.47.7.2186-2192.2003
  11. 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
  12. Chee, M. S., Lawrence, G. L. and Barrell, B. G. : Alpha-, betaand gammaherpesviruses encode a putative phosphotransferase. J. Gen. Virol. 70, 1151 (1989) https://doi.org/10.1099/0022-1317-70-5-1151
  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)
  15. Songyang, Z., Lu, K. P., Kwon, Y. T., Tsai, L.-H., Filhol, O., Cochet, C., Brickey, D. A., Soderling, T. R., Bartleson, C., Graves, D. J., Demaggio, A. J., Hoekstra, M. F., Blenis, J., Hunter, T. and Cantley, L. C. : A structural basis for substrate specificities of protein Ser/Thr kinases: primary sequence preference of casein kinases I and II, NIMA, phosphorylase kinase, calmodulin-dependent kinase II, CDK5, and Erk. Mol. Cell. Biol. 16, 6486 (1996) https://doi.org/10.1128/MCB.16.11.6486
  16. 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