Abstract
Three novel antimicrobial peptides (AMPs), named panurgines (PNGs), were isolated from the venom of the wild bee Panurgus calcaratus. The dodecapeptide of the sequence LNWGAILKHIIK-NH2 (PNG-1) belongs to the category of α-helical amphipathic AMPs. The other two cyclic peptides containing 25 amino acid residues and two intramolecular disulfide bridges of the pattern Cys8–Cys23 and Cys11–Cys19 have almost identical sequence established as LDVKKIICVACKIXPNPACKKICPK-OH (X=K, PNG-K and X=R, PNG-R). All three peptides exhibited antimicrobial activity against Gram-positive bacteria and Gram-negative bacteria, antifungal activity, and low hemolytic activity against human erythrocytes. We prepared a series of PNG-1 analogs to study the effects of cationicity, amphipathicity, and hydrophobicity on the biological activity. Several of them exhibited improved antimicrobial potency, particularly those with increased net positive charge. The linear analogs of PNG-K and PNG-R having all Cys residues substituted by α-amino butyric acid were inactive, thus indicating the importance of disulfide bridges for the antimicrobial activity. However, the linear PNG-K with all four cysteine residues unpaired, exhibited antimicrobial activity. PNG-1 and its analogs induced a significant leakage of fluorescent dye entrapped in bacterial membrane-mimicking large unilamellar vesicles as well as in vesicles mimicking eukaryotic cell membrane. On the other hand, PNG-K and PNG-R exhibited dye-leakage activity only from vesicles mimicking bacterial cell membrane.
Similar content being viewed by others
References
Amiche M, Galanth C (2011) Dermaseptins as models for the elucidation of membrane-acting helical amphipathic antimicrobial peptides. Curr Pharm Biotechnol 12:1184–1193
Argiolas A, Pisano JJ (1985) Bombolitins, a new class of mast cell degranulating peptides from the venom of the bumblebee Megabombus pennsylvanicus. J Biol Chem 260:1437–1444
Asthana N, Yadav SP, Ghosh JK (2004) Dissection of antimicrobial and toxic activity of melittin. J Biol Chem 279:55042–55050
Backlund B-M, Wikander G, Peeters T, Graslund A (1994) Induction of secondary structure in the peptide hormone motilin by interaction with phospholipid vesicles. Biochim Biophys Acta 1190:337–344
Baltzer SA, Brown MH (2011) Antimicrobial peptides—promising alternatives to conventional antibiotics. J Mol Microbiol Biotechnol 20:228–235
Brandenburg L-O, Merres J, Albrecht L-L, Varoga D, Pufe T (2012) Antimicrobial peptides: multifunctional drugs for different applications. Polymers 4:539–560
Čeřovský V, Slaninová J, Fučík V, Hulačová H, Borovičková L, Ježek R, Bednárová L (2008a) New potent antimicrobial peptides from the venom of Polistinae wasps and their analogs. Peptides 29:992–1003
Čeřovský V, Hovorka O, Cvačka J, Voburka Z, Bednárová L, Borovičková L, Slaninová J, Fučík V (2008b) Melectin: a novel antimicrobial peptide from the venom of the cleptoparasitic bee Melecta albifrons. ChemBioChem 9:2815–2821
Čeřovský V, Buděšínský M, Hovorka O, Cvačka J, Voburka Z, Slaninová J, Borovičková L, Fučík V, Bednárová L, Votruba I, Straka J (2009) Lasioglossins: three novel antimicrobial peptides from the venom of the eusocial bee Lasioglossum laticeps (Hymenoptera: Halictidae). ChemBioChem 10:2089–2099
Čeřovský V, Slaninová J, Fučík V, Monincová L, Bednárová L, Maloň P, Štokrová J (2011) Lucifensin, a novel insect defensin of medicinal maggots: synthesis and structural study. ChemBioChem 12:1352–1361
Chen Y, Guarnieri MT, Vasil AI, Vasil ML, Mant CT, Hodges RS (2007) Role of peptide hydrophobicity in the mechanism of action of α-helical antimicrobial peptides. Antimicrob Agents Chemother 51:1398–1406
Chou H-T, Wen H-W, Kuo T-Y, Lin C-C, Chen W-J (2010) Interaction of cationic antimicrobial peptides with phospholipid vesicles and their antibacterial activity. Peptides 31:1811–1820
Epand RM, Epand RF (2009) Domains in bacterial membranes and the action of antimicrobial agents. Mol BioSyst 5:580–587
Epand RM, Epand RF (2011) Bacterial membrane lipids in the action of antimicrobial targets. J Pept Sci 17:298–305
Epand RF, Savage PB, Epand RM (2007) Bacterial lipid composition and the antimicrobial efficacy of cationic steroid compounds (Ceragenins). Biochim Biophys Acta 1768:2500–2509
Giuliani A, Pirri G, Nicoletto SF (2007) Antimicrobial peptides: an overview of a promising class of therapeutics. Centr Eur J Biol 2:1–33
Huang Y, Huang J, Chen Y (2010) Alpha-helical cationic antimicrobial peptides: relationships of structure and function. Protein Cell 1:143–152
Jiang Z, Vasil AI, Hale JD, Hancock REW, Vasil ML, Hodges RS (2008) Effect of net charge and the number of positively charged residues on the biological activity of amphipathic α-helical cationic antimicrobial peptides. Biopolymers (Peptide Science) 90:369–383
Jones DT (1999) Protein secondary structure prediction based on position-specific scoring matrices. J Mol Biol 292:195–202
Kawai F, Shoda M, Harashima R, Sadaie Y, Hara H, Matsumoto K (2004) Cardiolipin domains in Bacillus subtilis Marburg membranes. J Bacteriol 186:1475–1483
Konno K, Hisada M, Fontana R, Lorenzi CCB, Naoki H, Itagaki Y, Miwa A, Kawai N, Nakata Y, Yasuhara T, Neto JR, de Azevedo WF Jr, Palma MS, Nakajima T (2001) Anoplin, a novel antimicrobial peptide from the venom of the solitary wasp Anoplius samariensis. Biochim Biophys Acta 1550:70–80
Konno K, Hisada M, Naoki H, Itagaki Y, Fontana R, Rangel M, Oliveira JS, Cabrera MPS, Neto JR, Hide I, Nakata Y, Yasuhara T, Nakajima T (2006) Eumenitin, a novel antimicrobial peptide from the venom of the solitary eumenine wasp Eumenes rubronotatus. Peptides 27:2624–2631
Konno K, Rangel M, Oliveira JS, dos Santos Cabrera MP, Fontana R, Hirata IY, Hide I, Nakata Y, Mori K, Kawano M, Fuchino H, Sekita S, Neto JR (2007) Decoralin, a novel linear cationic α-helical peptide from the venom of the solitary eumenine wasps Oreumenes decoratus. Peptides 28:2320–2327
Kuhn-Nentwig L (2003) Antimicrobial and cytolytic peptides of venomous arthropods. Cell Mol Life Sci 60:2651–2668
Labbé-Julié C, Granier C, Albericio F, Defendini M-L, Ceard B, Rochat H, Van Rietschoten J (1991) Binding and toxicity of apamin. Characterization of the active site. Eur J Biochem 196:639–645
Lohner K, Blondele SS (2005) Molecular mechanisms of membrane perturbation by antimicrobial peptides and the use of biophysical studies in the design of novel peptide antibiotics. Comb Chem High T Scr 8:241–256
Lyu PC, Sherman JC, Chen A, Kallenbach NR (1991) α-Helix stabilization by natural and unnatural amino acids with alkyl side chains. Proc Natl Acad Sci USA 88:5317–5320
Monincová L, Slaninová J, Voburka Z, Hovorka O, Fučík V, Borovičková L, Bednárová L, Buděšínský M, Straka J, Čeřovský V (2009) Novel biologically active peptides from the venom of the solitary bee Macropis fulvipes (Hymenoptera: Melittidae). In: Slaninová J (ed) Collection symposium series, institute of organic chemistry and biochemistry, vol 11. Academy of Sciences of the Czech Republic, Prague, pp 77–80
Monincová L, Buděšínský M, Slaninová J, Hovorka O, Cvačka J, Voburka Z, Fučík V, Borovičková L, Bednárová L, Straka J, Čeřovský V (2010) Novel antimicrobial peptides from the venom of the eusocial bee Halictus sexcinctus (Hymenoptera: Halictidae) and their analogs. Amino Acids 39:763–775
Monincová L, Slaninová J, Fučík V, Hovorka O, Voburka Z, Bednárová L, Maloň P, Štokrová J, Čeřovský V (2012) Lasiocepsin, a novel cyclic antimicrobial peptide from the venom of eusocial bee Lasioglossum laticeps (Hymenoptera: Halictidae). Amino Acids 43:751–761
Oyston PCF, Fox MA, Richards SJ, Clark GC (2009) Novel peptide therapeutics for treatment of infections. J Med Microb 58:977–987
Pazderková M, Kočišová E, Pazderka T, Maloň P, Kopecký Jr. V, Monincová L, Čeřovský V, Bednárová L (2012) Antimicrobial peptide from the eusocial bee Halictus sexcinctus. Interacting with model membranes. Spectroscopy Int J 27:497–502
Rohl CA, Baldwin RL (1998) Deciphering rules of helix stability in peptides. Methods Enzymol 295:1–26
Shailesh S, Neelam S, Sandeep K, Gupta GD (2009) Liposomes: a review. J Pharm Res 2:1163–1167
Teixeira V, Feio MJ, Bastos M (2012) Role of lipids in the interaction of antimicrobial peptides with membranes. Prog Lipid Res 51:149–177
Terashima H, Kojima S, Homma M (2008) Flagellar motility in bacteria: structure and function of flagellar motor. Int Rew Cell Moll Biol 270:39–85
Toke O (2005) Antimicrobial peptides: new candidates in the fight against bacterial infections. Biopolymers (Peptide Science) 80:717–735
Tossi A, Sandri L, Giangaspero A (2000) Amphipathic, α-helical antimicrobial peptides. Biopolymers (Peptide Science) 55:4–30
Vemuri S, Rhodes CT (1995) Preparation and characterization of liposomes as therapeutic delivery systems: a review. Pharm Acta Helvetica 70:95–111
Wang G, Li X, Wang Z (2009) APD2: the updated antimicrobial peptide database and its application in peptide design. Nucleic Acids Res 37:D933–D937
Whitmore L, Wallace BA (2008) Protein secondary structure analyses from circular dichroism spectroscopy: methods and reference databases. Biopolymers 89:392–400
Wieprecht T, Dathe M, Krause M, Beyermann M, Maloy WL, MacDonnald DL, Bienert M (1997) Modulation of membrane activity of amphipathic, antimicrobial peptides by slight modification of hydrophobic moment. FEBS Lett 417:135–140
Wimley WC, Hristova K (2011) Antimicrobial peptides: successes, challenges and unanswered questions. J Membr Biol 239:27–34
Yeaman MR, Yount NY (2003) Mechanisms of antimicrobial peptide action and resistance. Pharm Rev 55:27–55
Yeung ATY, Gellatly SL, Hancock REW (2011) Multifunctional cationic host defence peptides and their clinical applications. Cell Mol Life Sci 68:2161–2176
Zaiou M (2007) Multifunctional antimicrobial peptides: therapeutic targets in several human diseases. J Mol Med 85:317–329
Acknowledgments
This work was supported by the Grant Agency of the Charles University no. 645012, Czech Science Foundation, Grant no. 203/08/0536, and by research project RVO 61388963 of the Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic. We thank our technical assistants Mrs. Hana Hulačová and Mrs. Lenka Borovičková for their help with peptide synthesis. We also thank Gale A. Kirking at English Editorial Services, s.r.o. for assistance with the English.
Conflict of interest
The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Čujová, S., Slaninová, J., Monincová, L. et al. Panurgines, novel antimicrobial peptides from the venom of communal bee Panurgus calcaratus (Hymenoptera: Andrenidae). Amino Acids 45, 143–157 (2013). https://doi.org/10.1007/s00726-013-1482-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00726-013-1482-4