Abstract
Brevinin-1BYa (FLPILASLAAKFGPKLFCLVTKKC), first isolated from skin secretions of the frog Rana boylii, displays broad-spectrum antimicrobial activity and potent haemolytic activity. This study investigates the effects on conformation and biological activity of replacement of the intramolecular disulphide bridge in the peptide by a non-reducible dicarba bond. Dicarba-brevinin-1BYa was prepared by microwave irradiation of [Agl18,Agl24]-brevinin-1BYa (Agl = allylglycine) in the presence of a second generation Grubbs’ catalyst. Circular dichroism spectroscopy in 50% trifluoroethanol-water indicated that the degree of α-helicity of the dicarba derivative (22%) was less than that of brevinin-1BYa (27%) but comparable to that of the acyclic derivative [Ser18,Ser24]-brevinin-1BYa (23%). Dicarba-brevinin-1BYa showed a two-fold increase in potency against reference strains of Escherichia coli, Staphylococcus aureus, and Candida albicans compared with the native peptide and displayed potent bactericidal activity against clinical isolates of methicillin-resistant S. aureus (MRSA) and multidrug-resistant Acinetobacter baumannii (MIC in the range 1–8 μM). Compared with brevinin-1BYa and [Ser18,Ser24]-brevinin-1BYa, the dicarba derivative was associated with increased cytotoxicity against human erythrocytes (2.5-fold), MDA-MB-231 breast carcinoma cells (1.3-fold) and HepG2 hepatoma-derived cells (1.5-fold).
Similar content being viewed by others
Abbreviations
- CFU:
-
Colony forming units
- DC:
-
Dicarba
- DMF:
-
Dimethylformamide
- MDRAB:
-
Multidrug-resistant Acinetobacter baumannii
- MRSA:
-
Methicillin-resistant Staphylococcus aureus
- RCM:
-
Ring-closing metathesis
- TFA:
-
Trifluoroacetic acid
- TFE:
-
Trifluoroethanol
References
Auvynet C, Rosenstein Y (2009) Multifunctional host defense peptides: antimicrobial peptides, the small yet big players in innate and adaptive immunity. FEBS J 276:6497–6508
Barragán F, Moreno V, Marchán V (2009) Solid-phase synthesis and DNA binding studies of dichloroplatinum(ii) conjugates of dicarba analogues of octreotide as new anticancer drugs. Chem Commun 31:4705–4707
Bathgate RAD, Lin F, Hanson NF, Otvos L Jr, Guidolin A, Giannakis C, Bastiras S, Layfield S, Ferraro T, Ma S, Zhao C, Gundlach AL, Samuel CS, Tregear GW, Wade JD (2006) Relaxin-3: improved synthesis strategy and demonstration of its high-affinity interaction with the relaxin receptor LGR7 both in vitro and in vivo. Biochemistry 45:1043–1053
Bay J, Cederbaum AI (2006) Overexpression of CYP2E1 in mitochondria sensitizes HepG2 cells to the toxicity caused by depletion of glutathione. J Biol Chem 281:5128–5136
Clinical Laboratory and Standards Institute (2008a) Methods for dilution antimicrobial susceptibility tests for bacteria that grow aerobically. Approved standard M07-A8. CLSI, Wayne
Clinical Laboratory and Standards Institute (2008b) Reference method for broth dilution antifungal susceptibility testing of yeast. Approved standard M27-A3. CLSI, Wayne
Conlon JM (2008) Reflections on a systematic nomenclature for antimicrobial peptides from the skins of frogs of the family Ranidae. Peptides 29:1815–1819
Conlon JM (2011) The contribution of skin antimicrobial peptides to the system of innate immunity in anurans. Cell Tissue Res 343:201–212
Conlon JM, Sonnevend A, Patel M, Davidson C, Nielsen PF, Pál T, Rollins-Smith LA (2003) Isolation of peptides of the brevinin-1 family with potent candidacidal activity from the skin secretions of the frog Rana boylii. J Pept Res 62:207–213
Conlon JM, Al-Ghaferi N, Abraham B, Leprince J (2007) Strategies for transformation of naturally-occurring amphibian antimicrobial peptides into therapeutically valuable anti-infective agents. Methods 42:349–357
Conlon JM, Power GJ, Abdel-Wahab YHA, Flatt PR, Jiansheng H, Coquet L, Leprince J, Jouenne T, Vaudry H (2008) A potent, non-toxic insulin-releasing peptide isolated from an extract of the skin of the Asian frog, Hylarana güntheri (Anura:Ranidae). Regul Pept 151:153–159
Conlon JM, Ahmed E, Condamine E (2009a) Antimicrobial properties of brevinin-2-related peptide and its analogs: efficacy against multidrug-resistant Acinetobacter baumannii. Chem Biol Drug Des 74:488–493
Conlon JM, Kolodziejek J, Nowotny N (2009b) Antimicrobial peptides from the skins of North American frogs. Biochim Biophys Acta 1788:1556–1563
David MZ, Daum RS (2010) Community-associated methicillin-resistant Staphylococcus aureus: epidemiology and clinical consequences of an emerging epidemic. Clin Microbiol Rev 23:616–687
Deleo FR, Otto M, Kreiswirth BN, Chambers HF (2010) Community-associated methicillin-resistant Staphylococcus aureus. Lancet 375:1557–1568
Derksen DJ, Stymiest JL, Vederas JC (2006) Antimicrobial leucocin analogues with a disulfide bridge replaced by a carbocycle or by noncovalent interactions of allyl glycine residues. J Am Chem Soc 128:14252–14253
Diamond G, Beckloff N, Weinberg A, Kisich KO (2009) The roles of antimicrobial peptides in innate host defense. Curr Pharm Des 15:2377–2392
Forood B, Filiciano EJ, Niambiar KP (1993) Stabilization of α-helical structures in short peptides via end capping. Proc Natl Acad Sci USA 90:838–842
Frost DR (2010) Amphibian species of the world: an online reference. Version 5.4. American Museum of Natural History, New York, USA. http://www.research.amnh.org/herpetology/amphibia/index.php
Gootz TD, Marra A (2008) Acinetobacter baumannii: an emerging multidrug-resistant threat. Expert Rev Anti Infect Ther 6:309–325
Greenfield NJ (1996) Methods to estimate the conformation of proteins and polypeptides from circular dichroism data. Anal Biochem 235:1–10
Hossain MA, Rosengren KJ, Zhang S, Bathgate RA, Tregear GW, van Lierop BJ, Robinson AJ, Wade JD (2009) Solid phase synthesis and structural analysis of novel A-chain dicarba analogs of human relaxin-3 (INSL7) that exhibit full biological activity. Org Biomol Chem 7:1547–1553
Jacobsen Ø, Klaveness J, Rongved P (2010) Structural and pharmacological effects of ring-closing metathesis in peptides. Molecules 15:6638–6677
Kwon MY, Hong SY, Lee KH (1998) Structure-activity analysis of brevinin 1E amide, an antimicrobial peptide from Rana esculenta. Biochim Biophys Acta 1387:239–248
MacRaild CA, Illesinghe J, van Lierop BJ, Townsend AL, Chebib M, Livett BG, Robinson AJ, Norton RS (2009) Structure and activity of (2, 8)-dicarba-(3, 12)-cystino α-ImI, an α-conotoxin containing a nonreducible cystine analogue. J Med Chem 52:755–762
Maragakis LL, Perl TM (2008) Acinetobacter baumannii: epidemiology, antimicrobial resistance, and treatment options. Clin Infect Dis 46:1254–1263
Martinez M, Silley P (2010) Antimicrobial drug resistance. Handb Exp Pharmacol 199:227–264
Mechkarska M, Ahmed E, Coquet L, Leprince J, Jouenne T, Vaudry H, King JD, Conlon JM (2010) Antimicrobial peptides with therapeutic potential from skin secretions of the Marsabit clawed frog Xenopus borealis (Pipidae). Comp Biochem Physiol C Toxicol Pharmacol 152:467–472
Mookherjee N, Hancock RE (2007) Cationic host defence peptides: innate immune regulatory peptides as a novel approach for treating infections. Cell Mol Life Sci 64:922–933
Morikawa N, Hagiwara K, Nakajima T (1992) Brevinin-1 and -2, unique antimicrobial peptides from the skin of the frog, Rana brevipoda porsa. Biochem Biophys Res Commun 189:184–190
Muñoz V, Serrano L (1994) Elucidating the folding problem of helical peptides using empirical parameters. Nature Struct Biol 1:399–409
Nicolas P, El Amri C (2009) The dermaseptin superfamily: a gene-based combinatorial library of antimicrobial peptides. Biochim Biophys Acta 1788:1537–1550
Norrby SR, Nord CE, Finch R (2005) Lack of development of new antimicrobial drugs: a potential serious threat to public health. Lancet Infect Dis 5:115–119
Pál T, Abraham B, Sonnevend A, Jumaa P, Conlon JM (2006) Brevinin-1BYa: a naturally occurring peptide from frog skin with broad-spectrum antibacterial and antifungal properties. Int J Antimicrob Agents 27:525–529
Park SH, Kim HE, Kim CM, Yun HJ, Choi EC, Lee BJ (2002) Role of proline, cysteine and a disulphide bridge in the structure and activity of the anti-microbial peptide gaegurin 5. Biochem J 368:171–182
Perczel A, Fasman GD (1992) Quantitative analysis of cyclic beta-turn models. Protein Sci 1:378–395
Perez F, Hujer AM, Hujer KM, Decker BK, Rather PN, Bonomo RA (2007) Global challenge of multidrug-resistant Acinetobacter baumannii. Antimicrob Agents Chemother 51:3471–3484
Powers JP, Hancock RE (2003) The relationship between peptide structure and antibacterial activity. Peptides 24:1681–1691
Pukala TL, Bowie JH, Maselli VM, Musgrave IF, Tyler MJ (2006) Host-defence peptides from the glandular secretions of amphibians: structure and activity. Nat Prod Rep 23:368–393
Robinson AJ, Elaridi J, van Lierop BJ, Mujcinovic S, Jackson WR (2007) Microwave-assisted RCM for the synthesis of carbocyclic peptides. J Pept Sci 13:280–285
Robinson AJ, van Lierop BJ, Garland RD, Teoh E, Elaridi J, Illesinghe JP, Jackson WR (2009) Regioselective formation of interlocked dicarba-bridges in naturally occurring cyclic peptide toxins using olefin metathesis. Chem Commun 28:4293–4295
Stymiest JL, Mitchell BF, Wong S, Vederas JC (2003) Synthesis of biologically active dicarba analogues of the peptide hormone oxytocin using ring-closing metathesis. Org Lett 5:47–49
Tachi T, Epand RF, Epand RM, Matsuzaki K (2002) Position dependent hydrophobicity of the antimicrobial magainin peptide affects the mode of peptide-lipid interactions and selective toxicity. Biochemistry 41:10723–10731
Turton JF, Woodford N, Glover J, Yarde S, Kaufmann ME, Pitt TL (2006) Identification of Acinetobacter baumannii by detection of the blaOXA-51-like carbapenemase gene intrinsic to this species. J Clin Microbiol 44:2974–2976
Turton JF, Gabriel SN, Valderrey C, Kaufmann ME, Pitt TL (2007) Use of sequence-based typing and multiplex PCR to identify clonal lineages of outbreak strains of Acinetobacter baumannii. Clin Microbiol Infect 13:807–815
van Lierop BJ, Whelan AN, Andrikopoulos S, Mulder R, Jackson WR, Robinson AJ (2010) Methods for enhancing ring closing metathesis yield in peptides: synthesis of a dicarba human growth hormone fragment. Int J Pept Res Ther 16:133–144
Yeaman MR, Yount NY (2003) Mechanisms of antimicrobial peptide action and resistance. Pharmacol Rev 55:27–55
Zhang L, Falla TJ (2010) Potential therapeutic application of host defense peptides. Methods Mol Biol 618:303–327
Zhang S, Hughes RA, Bathgate RAD, Shabanpoora F, Hossain MA, Lin F, van Lierop BJ, Robinson AJ, Wade JD (2010) Role of the intra-A-chain disulfide bond of insulin-like peptide 3 in binding and activation of its receptor, RXFP2. Peptides 31:1730–1736
Acknowledgments
The authors thank Eman Ahmed, Milena Mechkarska, Kholoud Arafat and Annie Sunny (United Arab Emirates University) for technical assistance in this study. This work was supported by a Faculty Support Grant from the United Arab Emirates University.
Author information
Authors and Affiliations
Corresponding author
Additional information
Membrane-active peptides: 455th WE-Heraeus-Seminar and AMP 2010 Workshop.
Rights and permissions
About this article
Cite this article
Hossain, M.A., Guilhaudis, L., Sonnevend, A. et al. Synthesis, conformational analysis and biological properties of a dicarba derivative of the antimicrobial peptide, brevinin-1BYa. Eur Biophys J 40, 555–564 (2011). https://doi.org/10.1007/s00249-011-0679-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00249-011-0679-2