Identification and Characterization of Novel Antibacterial Peptides from Skin Secretions of Euphlyctis cyanophlyctis
- 275 Downloads
In this study, we extracted and purified antimicrobial peptides (AMPs) secreted from skin of Euphlyctis cyanophlyctis using reverse phase-high performance liquid chromatography. Three AMPs were isolated from skin secretions of this frog and sequenced using tandem mass spectrometry. The purified peptides were named buforin-EC (1875.05 ± 0.5 Da), cyanophlyctin (2347.50 ± 0.5 Da) and temporin-ECa (1013.33 ± 0.5 Da). Multiple alignments and homology search showed that buforin-EC, cyanophlyctin and temporin-ECa had a homology of 71.43, 47.1, and 69.23% to buforin II, brevinin-2EC, and temporin-1CSc, respectively. Antimicrobial tests demonstrated that our peptides have a great antimicrobial effect on both gram-positive and gram-negative bacteria. The results indicated that they have an overall minimum inhibitory concentration (MIC) below 13 μM against E. coli. No hemolysis was observed in around of their MIC values. In conclusion, skin secretions of E. cyanophlyctis contain a novel class of AMPs with the proper characteristics.
KeywordsAntimicrobial peptides Euphlyctis cyanophlyctis Hemolysis Minimum inhibitory concentration (MIC) RP-HPLC
This work was funded by grant number 87020301 from the Iran National Science Foundation (INSF).
- Asoodeh A, Zare Zardini H, Chamani J (2011) Identification and characterization of two novel antimicrobial peptides, temporin-Ra and temporin-Rb, from skin secretions of the marsh frog (Rana ridibunda). J Pept Sci. doi: 10.1002/psc.1409
- Conlon JM, Al-Dhaheri A, Al-Mutawa E, Al-Kharrge R, Ahmed E, Kolodziejek J, Nowotny N, Nielsen PF, Davidson C (2007) Peptide defenses of the Cascades frog Rana cascadae: implications for the evolutionary history of frogs of the Amerana species group. Peptides 28:1268–1274PubMedCrossRefGoogle Scholar
- Duellman WE (1999) Patterns of distribution of amphibians: a global perspective. The Johns Hopkins University Press, Baltimore, MDGoogle Scholar
- Ghavami S, Asoodeh A, Klonisch T, Halayko AJ, Kadkhoda K, Kroczak TJ, Gibson SB, Booy EP, Naderi-Manesh H, Los M (2008) Brevinin-2R1 semi-selectively kills cancer cells by a distinct mechanism, which involves the lysosomal-mitochondrial death pathway. J Cell Mol Med 12:1005–1022PubMedCrossRefGoogle Scholar
- Giacometti A, Cirioni O, Kamysz W, Silvestri C, Licci A, Riva A, Lukasiak J, Scalise G (2005) In vitro activity of amphibian peptides alone and in combination with antimicrobial agents against multidrug-resistant pathogens isolated from surgical wound infection. Peptides 26:2111–2116PubMedCrossRefGoogle Scholar
- Mehrnejad F, Naderi-Manesh H, Ranjbar B, Maroufi B, Asoodeh A, Doustdar F (2008) PCR-based gene synthesis, molecular cloning, high level expression, purification, and characterization of novel antimicrobial peptide, Brevinin-2R, in Escherichia coli. Appl Biochem Biotech 149:109–118CrossRefGoogle Scholar
- Minn I, Kim HS, Kim SC (1988) Antimicrobial peptides derived from pepsinogens in the stomach of the bullfrog, Rana catesbeiana. Biochim Biophys Acta 1407:31–39Google Scholar
- Ostorhazi ER, Sztodola F, Harmos A, Kovalszky F, Szabo I, Knappe D, Hoffmann D, Cassone R, Wade M, Bonomo JD, Otvos RA, Ir J (2010) Preclinical advantages of intramuscularly administered peptide A3-APO over existing therapies in Acinetobacter baumannii wound infections. J Antimicrob Chemother 65:2416–2422PubMedCrossRefGoogle Scholar