Advertisement

Biochemistry (Moscow)

, Volume 75, Issue 3, pp 336–341 | Cite as

Studies on amino acid replacement and inhibitory activity of a β-lactamase inhibitory peptide

  • Liping Xie
  • Mingfei Xu
  • Tao Yang
  • Chunbao Zhu
  • Baoquan Zhu
  • Youjia HuEmail author
Article

Abstract

An SHV β-lactamase gene was amplified from a β-lactam resistant Klebsiella pneumoniae K-71 genomic DNA. After expression and purification, we demonstrated that peptide P1 could inhibit the hydrolysis activity of both TEM-1 and SHV β-lactamase in vitro. Three mutations were introduced into P1 in which the first residue S was replaced by F, the 18th residue V was mutated to Y, and the 15th residue Y was substituted with A, C, G, and R to obtain the mutants of P1-A, P1- C, P1-G, and P1-R, respectively. The mutant peptides were purified and their inhibitory constants against TEM-1 and SHV β-lactamase were determined. All these β-lactamase inhibitory peptides could inhibit the activity of both β-lactamases, while the mutant peptides showed stronger inhibitory activities against TEM-1 β-lactamase than against SHV β-lactamase. Inhibition data suggested that P1-A improved the β-lactamase inhibitory activity by over 3-fold compare to P1. When P1-A was incubated with K. pneumoniae K-71 in Luria-Bertani medium containing ampicillin, it showed a much stronger growth of inhibition ratio over P1. This study gives us a good candidate for development of novel β-lactamase inhibitors.

Key words

β-lactamase inhibitory peptide β-lactamase amino acid replacement Klebsiella pneumoniae inhibitory constant 

Abbreviations

BLIP

β-lactamase inhibitory peptide

IPTG

isopropyl-β-D-thiogalactopyranoside

MIC

minimal inhibitory concentration

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Rasmussen, B. A., and Bush, K. (1997) Antimicrob. Agents Chemother., 41, 223–232.PubMedGoogle Scholar
  2. 2.
    Sykes, R. B., Bonner, D. P., Bush, K., and Georgopapadakou, N. H. (1982) Antimicrob. Agents Chemother., 21, 85–92.PubMedGoogle Scholar
  3. 3.
    Fisher, J., Charnas, R. L., and Knowles, J. R. (1978) Biochemistry, 17, 2180–2184.CrossRefPubMedGoogle Scholar
  4. 4.
    English, A. R., Retsema, J. A., Girard, A. E., Lynch, J. E., and Barth, W. E. (1978) Antimicrob. Agents Chemother., 14, 414–419.PubMedGoogle Scholar
  5. 5.
    Micetich, R. G., Maiti, S. N., Spevak, P., Hall, T. W., Yamabe, S., Ishida, N., Tanaka, M., Yamazaki, T., Nakai, A., and Ogawa, K. (1987) J. Med. Chem., 30, 1469–1474.CrossRefPubMedGoogle Scholar
  6. 6.
    Doran, J. L., Leskiw, B. K., Aippersbach, S., and Jensen, S. E. (1990) J. Bacteriol., 172, 4909–4918.PubMedGoogle Scholar
  7. 7.
    Rudgers, G. W., Huang, W., and Palzkill, T. (2001) Antimicrob. Agents Chemother., 45, 3279–3286.CrossRefPubMedGoogle Scholar
  8. 8.
    Sun, W., Hu, Y., Gong, J., Zhu, C., and Zhu, B. (2005) Biochemistry (Moscow), 70, 753–760.CrossRefGoogle Scholar
  9. 9.
    Wang, J., Palzkill, T., and Chow, D. C. (2009) J. Biol. Chem., 284, 595–609.CrossRefPubMedGoogle Scholar
  10. 10.
    Zhang, Z., and Palzkill, T. (2003) J. Biol. Chem., 278, 45706–45712.CrossRefPubMedGoogle Scholar
  11. 11.
    Pospiech, A., and Neumann, B. (1995) Trends Genet., 11, 217–218.CrossRefPubMedGoogle Scholar
  12. 12.
    Petrosino, J., Rudgers, G., Gilbert, H., and Palzkill, T. (1999) J. Biol. Chem., 274, 2394–2400.CrossRefPubMedGoogle Scholar
  13. 13.
    Cornish-Bowden, A. (1974) Biochem. J., 137, 143–144.PubMedGoogle Scholar
  14. 14.
    Dixon, M. (1953) Biochem. J., 55, 170–171.PubMedGoogle Scholar
  15. 15.
    Ness, S., Martin, R., Kindler, A. M., Paetzel, M., Gold, M., Jensen, S. E., Jones, J. B., and Strynadka, N. C. (2000) Biochemistry, 39, 5312–5321.CrossRefPubMedGoogle Scholar
  16. 16.
    Reichmann, D., Rahat, O., Albeck, S., Meged, R., Dym, O., and Schreiber, G. (2005) Proc. Natl. Acad. Sci. USA, 102, 57–62.CrossRefPubMedGoogle Scholar
  17. 17.
    Thai, W., Paradkar, A. S., and Jensen, S. E. (2001) Microbiology, 147, 325–335.PubMedGoogle Scholar
  18. 18.
    Strynadka, N. C., Jensen, S. E., Alzari, P. M., and James, M. N. (1996) Nature Struct. Biol., 3, 290–297.CrossRefPubMedGoogle Scholar
  19. 19.
    Zhang, Z., and Palzkill, T. (2004) J. Biol. Chem., 279, 42860–42866.CrossRefPubMedGoogle Scholar
  20. 20.
    Albeck, S., and Schreiber, G. (1999) Biochemistry, 38, 11–21.CrossRefPubMedGoogle Scholar
  21. 21.
    Reynolds, K. A., Thomson, J. M., Corbett, K. D., Bethel, C. R., Berger, J. M., Kirsch, J. F., Bonomo, R. A., and Handel, T. M. (2006) J. Biol. Chem., 281, 26745–267453.CrossRefPubMedGoogle Scholar
  22. 22.
    Selzer, T., Albeck, S., and Schreiber, G. (2000) Nature Struct. Biol., 7, 537–541.CrossRefPubMedGoogle Scholar
  23. 23.
    Yao, J., and Asakura, T. (2003) J. Biochem., 133, 147–154.CrossRefPubMedGoogle Scholar
  24. 24.
    Rice, L. B., Carias, L. L., Hujer, A. M., Bonafede, M., Hutton, R., Hoyen, C., and Bonomo, R. A. (2000) Antimicrob. Agents Chemother., 44, 362–367.CrossRefPubMedGoogle Scholar
  25. 25.
    DiPersio, J. R., Deshpande, L. M., Biedenbach, D. J., Toleman, M. A., Walsh, T. R., and Jones, R. N. (2005) Diagn. Microbiol. Infect. Dis., 51, 1–7.CrossRefPubMedGoogle Scholar

Copyright information

© Pleiades Publishing, Ltd. 2010

Authors and Affiliations

  • Liping Xie
    • 1
  • Mingfei Xu
    • 1
  • Tao Yang
    • 1
  • Chunbao Zhu
    • 1
  • Baoquan Zhu
    • 1
  • Youjia Hu
    • 1
    Email author
  1. 1.State Key Laboratory of New Drug and Pharmaceutical ProcessShanghai Institute of Pharmaceutical IndustryShanghaiChina

Personalised recommendations