• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biochemjBJ Latest papers and much more!
Biochem J. May 1, 2000; 347(Pt 3): 633–641.
PMCID: PMC1220998

Large-scale synthesis and functional elements for the antimicrobial activity of defensins.

Abstract

Human neutrophil defensins, and their analogues incorporating anionic, hydrophobic or cationic residues at the N- and C-termini, were synthesized by solid-phase procedures. The synthetic defensins were examined for their microbicidal activity against Candida albicans, two Gram-negative bacteria (Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis) and two Gram-positive bacteria (Streptococcus gordonii and Streptococcus mutans). The human neutrophil peptide 1 (HNP1) and HNP2 were found to be potent candidacidal agents. HNP3, which differs by one amino acid at the N-terminus of its sequence, was totally inactive. The Gram-negative bacteria A. actinomycetemcomitans and P. gingivalis and the Gram-positive bacteria S. gordonii and S. mutans were insensitive to human defensins. However, the insertion of two basic residues, such as arginine, at both the N-terminus and the C-terminus of HNP2 significantly enhanced antifungal and antibacterial activity. The addition of anionic residues, such as aspartic acid, at the N- and C-termini rendered the molecule totally inactive. The presence of two hydrophobic amino acids, such as valine, at the N-terminus of HNP2 and of two basic arginine residues at its C-terminus resulted in molecules that were optimally active against these oral pathogens. The results suggest that the N- and C-terminal residues in defensin peptides are the crucial functional elements that determine their microbicidal potency. The three-dimensional structure of all defensins constitutes the same amphiphilic beta-sheet structure, with the polar face formed by the N- and C-terminal residues playing an important role in defining microbicidal potency and the antimicrobial spectrum. The enhanced microbicidal activity observed for defensin peptides with two basic residues at both the N- and C-termini could be due to optimization of the amphiphilicity of the structure, which could facilitate specific interactions with the microbial membranes.

Full Text

The Full Text of this article is available as a PDF (210K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Patterson-Delafield J, Martinez RJ, Lehrer RI. Microbicidal cationic proteins in rabbit alveolar macrophages: a potential host defense mechanism. Infect Immun. 1980 Oct;30(1):180–192. [PMC free article] [PubMed]
  • Selsted ME, Brown DM, DeLange RJ, Lehrer RI. Primary structures of MCP-1 and MCP-2, natural peptide antibiotics of rabbit lung macrophages. J Biol Chem. 1983 Dec 10;258(23):14485–14489. [PubMed]
  • Lehrer RI, Lichtenstein AK, Ganz T. Defensins: antimicrobial and cytotoxic peptides of mammalian cells. Annu Rev Immunol. 1993;11:105–128. [PubMed]
  • García-Olmedo F, Molina A, Alamillo JM, Rodríguez-Palenzuéla P. Plant defense peptides. Biopolymers. 1998;47(6):479–491. [PubMed]
  • Diamond G, Zasloff M, Eck H, Brasseur M, Maloy WL, Bevins CL. Tracheal antimicrobial peptide, a cysteine-rich peptide from mammalian tracheal mucosa: peptide isolation and cloning of a cDNA. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3952–3956. [PMC free article] [PubMed]
  • Bensch KW, Raida M, Mägert HJ, Schulz-Knappe P, Forssmann WG. hBD-1: a novel beta-defensin from human plasma. FEBS Lett. 1995 Jul 17;368(2):331–335. [PubMed]
  • Schonwetter BS, Stolzenberg ED, Zasloff MA. Epithelial antibiotics induced at sites of inflammation. Science. 1995 Mar 17;267(5204):1645–1648. [PubMed]
  • Ouellette AJ, Selsted ME. Paneth cell defensins: endogenous peptide components of intestinal host defense. FASEB J. 1996 Sep;10(11):1280–1289. [PubMed]
  • Zhao C, Wang I, Lehrer RI. Widespread expression of beta-defensin hBD-1 in human secretory glands and epithelial cells. FEBS Lett. 1996 Nov 4;396(2-3):319–322. [PubMed]
  • Harder J, Bartels J, Christophers E, Schröder JM. A peptide antibiotic from human skin. Nature. 1997 Jun 26;387(6636):861–861. [PubMed]
  • McCray PB, Jr, Bentley L. Human airway epithelia express a beta-defensin. Am J Respir Cell Mol Biol. 1997 Mar;16(3):343–349. [PubMed]
  • Haynes RJ, Tighe PJ, Dua HS. Antimicrobial defensin peptides of the human ocular surface. Br J Ophthalmol. 1999 Jun;83(6):737–741. [PMC free article] [PubMed]
  • Mathews M, Jia HP, Guthmiller JM, Losh G, Graham S, Johnson GK, Tack BF, McCray PB., Jr Production of beta-defensin antimicrobial peptides by the oral mucosa and salivary glands. Infect Immun. 1999 Jun;67(6):2740–2745. [PMC free article] [PubMed]
  • Lehrer RI, Ganz T, Selsted ME. Oxygen-independent bactericidal systems. Mechanisms and disorders. Hematol Oncol Clin North Am. 1988 Mar;2(1):159–169. [PubMed]
  • Kagan BL, Ganz T, Lehrer RI. Defensins: a family of antimicrobial and cytotoxic peptides. Toxicology. 1994 Feb 28;87(1-3):131–149. [PubMed]
  • Diamond G, Zasloff M, Eck H, Brasseur M, Maloy WL, Bevins CL. Tracheal antimicrobial peptide, a cysteine-rich peptide from mammalian tracheal mucosa: peptide isolation and cloning of a cDNA. Proc Natl Acad Sci U S A. 1991 May 1;88(9):3952–3956. [PMC free article] [PubMed]
  • Weinberg A, Krisanaprakornkit S, Dale BA. Epithelial antimicrobial peptides: review and significance for oral applications. Crit Rev Oral Biol Med. 1998;9(4):399–414. [PubMed]
  • Selsted ME, Szklarek D, Lehrer RI. Purification and antibacterial activity of antimicrobial peptides of rabbit granulocytes. Infect Immun. 1984 Jul;45(1):150–154. [PMC free article] [PubMed]
  • Lehrer RI, Barton A, Daher KA, Harwig SS, Ganz T, Selsted ME. Interaction of human defensins with Escherichia coli. Mechanism of bactericidal activity. J Clin Invest. 1989 Aug;84(2):553–561. [PMC free article] [PubMed]
  • Cociancich S, Ghazi A, Hetru C, Hoffmann JA, Letellier L. Insect defensin, an inducible antibacterial peptide, forms voltage-dependent channels in Micrococcus luteus. J Biol Chem. 1993 Sep 15;268(26):19239–19245. [PubMed]
  • Hill CP, Yee J, Selsted ME, Eisenberg D. Crystal structure of defensin HNP-3, an amphiphilic dimer: mechanisms of membrane permeabilization. Science. 1991 Mar 22;251(5000):1481–1485. [PubMed]
  • Stanfield RL, Westbrook EM, Selsted ME. Characterization of two crystal forms of human defensin neutrophil cationic peptide 1, a naturally occurring antimicrobial peptide of leukocytes. J Biol Chem. 1988 Apr 25;263(12):5933–5935. [PubMed]
  • Levy RM, Bassolino DA, Kitchen DB, Pardi A. Solution structures of proteins from NMR data and modeling: alternative folds for neutrophil peptide 5. Biochemistry. 1989 Nov 28;28(24):9361–9372. [PubMed]
  • Zhang XL, Selsted ME, Pardi A. NMR studies of defensin antimicrobial peptides. 1. Resonance assignment and secondary structure determination of rabbit NP-2 and human HNP-1. Biochemistry. 1992 Nov 24;31(46):11348–11356. [PubMed]
  • Pardi A, Zhang XL, Selsted ME, Skalicky JJ, Yip PF. NMR studies of defensin antimicrobial peptides. 2. Three-dimensional structures of rabbit NP-2 and human HNP-1. Biochemistry. 1992 Nov 24;31(46):11357–11364. [PubMed]
  • Skalicky JJ, Selsted ME, Pardi A. Structure and dynamics of the neutrophil defensins NP-2, NP-5, and HNP-1: NMR studies of amide hydrogen exchange kinetics. Proteins. 1994 Sep;20(1):52–67. [PubMed]
  • Xiong YQ, Kupferwasser LI, Zack PM, Bayer AS. Comparative efficacies of liposomal amikacin (MiKasome) plus oxacillin versus conventional amikacin plus oxacillin in experimental endocarditis induced by Staphylococcus aureus: microbiological and echocardiographic analyses. Antimicrob Agents Chemother. 1999 Jul;43(7):1737–1742. [PMC free article] [PubMed]
  • Lehrer RI, Ganz T, Szklarek D, Selsted ME. Modulation of the in vitro candidacidal activity of human neutrophil defensins by target cell metabolism and divalent cations. J Clin Invest. 1988 Jun;81(6):1829–1835. [PMC free article] [PubMed]
  • Miyasaki KT, Bodeau AL, Ganz T, Selsted ME, Lehrer RI. In vitro sensitivity of oral, gram-negative, facultative bacteria to the bactericidal activity of human neutrophil defensins. Infect Immun. 1990 Dec;58(12):3934–3940. [PMC free article] [PubMed]
  • Kohashi O, Ono T, Ohki K, Soejima T, Moriya T, Umeda A, Meno Y, Amako K, Funakosi S, Masuda M, et al. Bactericidal activities of rat defensins and synthetic rabbit defensins on Staphylococci, Klebsiella pneumoniae (Chedid, 277, and 8N3), Pseudomonas aeruginosa (mucoid and nonmucoid strains), Salmonella typhimurium (Ra, Rc, Rd, and Re of LPS mutants) and Escherichia coli. Microbiol Immunol. 1992;36(4):369–380. [PubMed]
  • Kaiser E, Colescott RL, Bossinger CD, Cook PI. Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides. Anal Biochem. 1970 Apr;34(2):595–598. [PubMed]
  • Sundstrom PM, Nichols EJ, Kenny GE. Antigenic differences between mannoproteins of germ tubes and blastospores of Candida albicans. Infect Immun. 1987 Mar;55(3):616–620. [PMC free article] [PubMed]
  • Raj PA, Edgerton M, Levine MJ. Salivary histatin 5: dependence of sequence, chain length, and helical conformation for candidacidal activity. J Biol Chem. 1990 Mar 5;265(7):3898–3905. [PubMed]
  • Antonyraj KJ, Karunakaran T, Raj PA. Bactericidal activity and poly-L-proline II conformation of the tandem repeat sequence of human salivary mucin glycoprotein (MG2). Arch Biochem Biophys. 1998 Aug 15;356(2):197–206. [PubMed]
  • Dentino AR, Raj PA, Bhandary KK, Wilson ME, Levine MJ. Role of peptide backbone conformation on biological activity of chemotactic peptides. J Biol Chem. 1991 Oct 5;266(28):18460–18468. [PubMed]
  • Ganz T, Selsted ME, Szklarek D, Harwig SS, Daher K, Bainton DF, Lehrer RI. Defensins. Natural peptide antibiotics of human neutrophils. J Clin Invest. 1985 Oct;76(4):1427–1435. [PMC free article] [PubMed]
  • Kahn PC. The interpretation of near-ultraviolet circular dichroism. Methods Enzymol. 1979;61:339–378. [PubMed]
  • Lauth X, Nesin A, Briand JP, Roussel JP, Hetru C. Isolation, characterization and chemical synthesis of a new insect defensin from Chironomus plumosus (Diptera). Insect Biochem Mol Biol. 1998 Dec;28(12):1059–1066. [PubMed]
  • Rao AG, Rood T, Maddox J, Duvick J. Synthesis and characterization of defensin NP-1. Int J Pept Protein Res. 1992 Dec;40(6):507–514. [PubMed]
  • Wu M, Hancock RE. Interaction of the cyclic antimicrobial cationic peptide bactenecin with the outer and cytoplasmic membrane. J Biol Chem. 1999 Jan 1;274(1):29–35. [PubMed]
  • Wu M, Maier E, Benz R, Hancock RE. Mechanism of interaction of different classes of cationic antimicrobial peptides with planar bilayers and with the cytoplasmic membrane of Escherichia coli. Biochemistry. 1999 Jun 1;38(22):7235–7242. [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...