Identification of crucial residues for the antibacterial activity of the proline-rich peptide, pyrrhocoricin

Goran Kragol, Ralf Hoffmann, Michael A. Chattergoon, Sandor Lovas, Mare Cudic, Philippe Bulet, Barry A. Condie, K. Johan Rosengren, Luis J. Montaner, Laszlo Otvos

Research output: Contribution to journalArticle

Abstract

Members of the proline-rich antibacterial peptide family, pyrrhocoricin, apidaecin and drosocin appear to kill responsive bacterial species by binding to the multihelical lid region of the bacterial DnaK protein. Pyrrhocoricin, the most potent among these peptides, is nontoxic to healthy mice, and can protect these animals from bacterial challenge. A structure-antibacterial activity study of pyrrhocoricin against Escherichia coli and Agrobacterium tumefaciens identified the N-terminal half, residues 2-10, the region responsible for inhibition of the ATPase activity, as the fragment that contains the active segment. While fluorescein-labeled versions of the native peptides entered E. coli cells, deletion of the C-terminal half of pyrrhocoricin significantly reduced the peptide's ability to enter bacterial or mammalian cells. These findings highlighted pyrrhocoricin's suitability for combating intracellular pathogens and raised the possibility that the proline-rich antibacterial peptides can deliver drug leads into mammalian cells. By observing strong relationships between the binding to a synthetic fragment of the target protein and antibacterial activities of pyrrhocoricin analogs modified at strategic positions, we further verified that DnaK was the bacterial target macromolecule. In addition, the antimicrobial activity spectrum of native pyrrhocoricin against 11 bacterial and fungal strains and the binding of labeled pyrrhocoricin to synthetic DnaK D-E helix fragments of the appropriate species could be correlated. Mutational analysis on a synthetic E. coli DnaK fragment identified a possible binding surface for pyrrhocoricin.

Original languageEnglish (US)
Pages (from-to)4226-4237
Number of pages12
JournalEuropean Journal of Biochemistry
Volume269
Issue number17
DOIs
StatePublished - Sep 25 2002
Externally publishedYes

Keywords

  • Antimicrobial peptides
  • Cell penetration
  • Heat shock proteins
  • Mutational analysis
  • Pharmacophore

ASJC Scopus subject areas

  • Biochemistry

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  • Cite this

    Kragol, G., Hoffmann, R., Chattergoon, M. A., Lovas, S., Cudic, M., Bulet, P., Condie, B. A., Rosengren, K. J., Montaner, L. J., & Otvos, L. (2002). Identification of crucial residues for the antibacterial activity of the proline-rich peptide, pyrrhocoricin. European Journal of Biochemistry, 269(17), 4226-4237. https://doi.org/10.1046/j.1432-1033.2002.03119.x