Amino Acids

, Volume 40, Issue 1, pp 61–68 | Cite as

Amino acid substitutions in an alpha-helical antimicrobial arachnid peptide affect its chemical properties and biological activity towards pathogenic bacteria but improves its therapeutic index

  • A. Rodríguez
  • E. Villegas
  • H. Satake
  • L. D. Possani
  • Gerardo Corzo
Original Article

Abstract

Four variants of the highly hemolytic antimicrobial peptide Pin2 were chemically synthesized with the aim to investigate the role of the proline residue in this peptide, by replacing it with the motif glycine-valine-glycine [GVG], which was found to confer low hemolytic activity in a spider antimicrobial peptide. The proline residue in position 14 of Pin2 was substituted by [V], [GV], [VG] and [GVG]. Only the peptide variant with the proline substituted for [GVG] was less hemolytic compared to that of all other variants. The peptide variant [GVG] kept its antimicrobial activity in Muller–Hilton agar diffusion assays, whereas the other three variants were less effective. However, all Pin2 antimicrobial peptide variants, were active when challenged against a Gram-positive bacteria in Muller–Hilton broth assays suggesting that chemical properties of the antimicrobial peptides such as hydrophobicity is an important indication for antimicrobial activity in semi-solid environments.

Keywords

Antimicrobial peptide Arachnid, pandinin 2 Peptide synthesis S. aureus 

Abbreviations

CD

Circular dichroism

OD

Optical density

Oxki2

Oxypinin 2

Pin2

Pandinin 2

TFA

Trifluoroacetic acid

TFE

Trifluoroethanol

MIC

Minimum inhibitory concentration

References

  1. Amiche M, Seon AA, Wroblewski H, Nicolas P (2000) Isolation of dermatoxin from frog skin, an antibacterial peptide encoded by a novel member of the dermaseptin genes family. Eur J Biochem 267:4583–4592CrossRefPubMedGoogle Scholar
  2. Andreu D, Rivas L (1998) Animal antimicrobial peptides: an overview. Biopolymers 47:415–433CrossRefPubMedGoogle Scholar
  3. Batista CVF, Rosendo da Silva L, Sebben A, Scaloni A, Ferrara L, Paiva GR, Olamendi-Portugal T, Possani LD, Bloch C Jr (1999) Antimicrobial peptides from the Brazilian frog Phyllomedusa distincta. Peptides 20:679–686CrossRefPubMedGoogle Scholar
  4. Bazzo R, Tappin MJ, Pastore A, Harvey TS, Carver JA, Campbell ID (1988) The structure of melittin. A 1H-NMR study in methanol. Eur J Biochem 173:139–146CrossRefPubMedGoogle Scholar
  5. Bohm G, Muhr R, Jaenicke R (1992) Quantitative analysis of protein far UV circular dichroism spectra by neural networks. Protein Eng 5:191–195CrossRefPubMedGoogle Scholar
  6. Bywater RP, Thomas D, Vriend G (2001) A sequence and structural study of transmembrane helices. J Comput Aided Mol Des 15:533–552CrossRefPubMedGoogle Scholar
  7. Chandrapati S, O’Sullivan DJ (1998) Procedure for quantifiable assessment of nutritional parameters influencing Nisin production by Lactococcus lactis subsp lactis. J Biotechnol 63:229–233CrossRefPubMedGoogle Scholar
  8. Cordes FS, Bright JN, Sansom MSP (2002) Proline-induced distortions of transmembrane helices. J Mol Biol 323:951–960CrossRefPubMedGoogle Scholar
  9. Corzo G, Escoubas P, Villegas E, Barnham KJ, He W, Norton RS, Nakajima T (2001) Characterization of unique amphipathic antimicrobial peptides from venom of the scorpion Pandinus imperator. Biochem J 359:35–45CrossRefPubMedGoogle Scholar
  10. Corzo G, Villegas E, Gomez-Lagunas F, Possani LD, Belokoneva OS, Nakajima T (2002) Oxyopinins, large amphipathic peptides isolated from the venom of the wolf spider Oxyopes kitabensis with cytolytic properties and positive insecticidal cooperativity with spider neurotoxins. J Biol Chem 277:23627–23637CrossRefPubMedGoogle Scholar
  11. Dathe M, Kaduk C, Tachikawa E, Melzig MF, Wenschuh H, Bienert M (1998) Proline at position 14 of alamethicin is essential for hemolytic activity, catecholamine secretion from chromaffin cells and enhanced metabolic activity in endothelial cells. Biochim Biophys Acta Biomembr 1370:175–183CrossRefGoogle Scholar
  12. Dempsey CE, Bazzo R, Harvey TS, Syperek I, Boheim G, Campbell ID (1991) Contribution of proline-14 to the structure and actions of melittin. FEBS Lett 281:240–244CrossRefPubMedGoogle Scholar
  13. Holak TA, Engstrom A, Kraulis PJ, Lindeberg G, Bennich H, Jones TA, Gronenborn AM, Clore GM (1988) The solution conformation of the antibacterial peptide cecropin A: a nuclear magnetic resonance and dynamical simulated annealing study. Biochemistry 27:7620–7629CrossRefPubMedGoogle Scholar
  14. Iwai H, Nakajima Y, Natori S, Arata Y, Shimada I (1993) Solution conformation of an antibacterial peptide, sarcotoxin IA, as determined by 1H-NMR. Eur J Biochem 217:639–644CrossRefPubMedGoogle Scholar
  15. Kyte J, Doolittle RF (1982) A simple method for displaying the hydropathic character of a protein. J Mol Biol 157:105–132CrossRefPubMedGoogle Scholar
  16. Mor A, Nguyen VH, Delfour A, Migliore-Samour D, Nicolas P (1991) Isolation, amino acid sequence, and synthesis of dermaseptin, a novel antimicrobial peptide of amphibian skin. Biochemistry 30:8824–8830CrossRefPubMedGoogle Scholar
  17. Oren Z, Shai Y (1997) Selective lysis of bacteria but not mammalian cells by diastereomers of melittin: structure–function study. Biochemistry 36:1826–1835CrossRefPubMedGoogle Scholar
  18. Orivel J, Redeker V, Le Caer JP, Krier F, Revol-Junelles AM, Longeon A, Chaffotte A, Dejean A, Rossier J (2001) Ponericins, new antibacterial and insecticidal peptides from the venom of the ant Pachycondyla goeldii. J Biol Chem 276:17823–17829CrossRefPubMedGoogle Scholar
  19. Sansom MSP (1992) Proline residues in transmembrane helices of channel and transport proteins: a molecular modelling study. Protein Eng 5:53–60CrossRefPubMedGoogle Scholar
  20. Shin SY, Kang JH, Lee DG, Jang SY, Seo MY (1999) Influences of hinge region of a synthetic antimicrobial peptide, cecropin A(1–13)-melittin(1–13) hybrid on antibiotic activity. Bull Korean Chem Soc 20:1078–1084Google Scholar
  21. Thennarasu S, Nagaraj R (1996) Specific antimicrobial and hemolytic activities of 18-residue peptides derived from the amino terminal region of the toxin pardaxin. Protein Eng 9:1219–1224CrossRefPubMedGoogle Scholar
  22. Wong H, Bowie JH, Carver JA (1997) The solution structure and activity of caerin 1.1, an antimicrobial peptide from the Australian green tree frog, Litoria splendida. Eur J Biochem 247:545–557CrossRefPubMedGoogle Scholar
  23. Zidovetzki R, Rost B, Armstrong DL, Pecht I (2003) Transmembrane domains in the functions of Fc receptors. Biophys Chem 1000:555–575CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  • A. Rodríguez
    • 1
  • E. Villegas
    • 2
  • H. Satake
    • 3
  • L. D. Possani
    • 1
  • Gerardo Corzo
    • 1
  1. 1.Departamento de Medicina Molecular y Bioprocesos, Instituto de BiotecnologíaUniversidad Nacional Autónoma de México, UNAMCuernavacaMexico
  2. 2.Centro de Investigación en BiotecnologíaUniversidad Autónoma del Estado de Morelos, Av. Universidad 1001CuernavacaMexico
  3. 3.Suntory Institute for Bioorganic ResearchShimamoto-Cho, Mishima-GunJapan

Personalised recommendations