Model peptides mimic the structure and function of the N-terminus of the pore-forming toxin sticholysin II

Fábio Casallanovo, Felipe J F De Oliveira, Fernando C. De Souza, Uris Ros, Yohanka Martínez, David Penton, Mayra Tejuca, Diana Martínez, Fabiola Pazos, Thelma A. Pertinhez, Alberto Spisni, Eduardo M. Cilli, Maria E. Lanio, Carlos Alvarez, Shirley Schreier

Research output: Contribution to journalArticle

Abstract

To investigate the role of the N-terminal region in the lytic mechanism of the pore-forming toxin sticholysin II (St II), we studied the conformational and functional properties of peptides encompassing the first 30 residues of the protein. Peptides containing residues 1-30 (P1-30) and 11-30 (P11-30) were synthesized and their conformational properties were examined in aqueous solution as a function of peptide concentration, pH, ionic strength, and addition of the secondary structure-inducing solvent trifluoroethanol (TFE). CD spectra showed that increasing concentration, pH, and ionic strength led to aggregation of P1-30; as a consequence, the peptide acquired β-sheet conformation. In contrast, P11-30 exhibited practically no conformational changes under the same conditions, remaining essentially structureless. Moreover, this peptide did not undergo aggregation. These differences clearly point to the modulating effect of the first 10 hydrophobic residues on the peptides aggregation and conformational properties. In TFE both the first ten hydrophobic peptides acquired α-helical conformation, albeit to a different extent, P11-30 displayed lower α-helical content. P1-30 presented a larger fraction of residues in α-helical conformation in TFE than that found in St II's crystal structure for that portion of the protein. Since TFE mimics the membrane environment, such increase in helical content could also occur upon toxin binding to membranes and represent a step in the mechanism of pore formation. The peptides conformational properties correlated well with their functional behavior. Thus, P1-30 exhibited much higher hemolytic activity than P11-30. In addition, P11-30 was able to block the toxin s hemolytic activity. The size of pores formed in red blood cells by P1-30 was estimated by measuring the permeability to PEGs of different molecular mass. The pore radius (0.95 ± 0.01 nm) was very similar to that of the pore formed by the toxin. The results demonstrate that the synthetic peptide P1-30 is a good model of St II conformation and function and emphasize the contribution of the toxin's N-terminal region, and, in particular, the hydrophobic residues 1-10 to pore formation.

Original languageEnglish
Pages (from-to)169-180
Number of pages12
JournalBiopolymers
Volume84
Issue number2
DOIs
Publication statusPublished - 2006

Keywords

  • Actinoporin
  • Circular dichroism
  • Hemolytic peptide
  • Pore-forming toxin
  • Sticholysin II

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics

Fingerprint Dive into the research topics of 'Model peptides mimic the structure and function of the N-terminus of the pore-forming toxin sticholysin II'. Together they form a unique fingerprint.

  • Cite this

    Casallanovo, F., De Oliveira, F. J. F., De Souza, F. C., Ros, U., Martínez, Y., Penton, D., Tejuca, M., Martínez, D., Pazos, F., Pertinhez, T. A., Spisni, A., Cilli, E. M., Lanio, M. E., Alvarez, C., & Schreier, S. (2006). Model peptides mimic the structure and function of the N-terminus of the pore-forming toxin sticholysin II. Biopolymers, 84(2), 169-180. https://doi.org/10.1002/bip.20374