This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Chapple, D. S.
Right arrow Articles by Evans, R. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Chapple, D. S.
Right arrow Articles by Evans, R. W.

 Previous Article  |  Next Article 

Infect Immun, June 1998, p. 2434-2440, Vol. 66, No. 6
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Structure-Function Relationship of Antibacterial Synthetic Peptides Homologous to a Helical Surface Region on Human Lactoferrin against Escherichia coli Serotype O111

Daniel S. Chapple,1,2,3 David J. Mason,3 Christopher L. Joannou,1 Edward W. Odell,2 Vanya Gant,3 and Robert W. Evans1,*

Metalloprotein Research Group, Division of Biochemistry and Molecular Biology,1 and Department of Oral Medicine and Pathology,2 UMDS, Guy's Hospital, London SE1 9RT, and Infection and Immunity Laboratory, Division of Infection, UMDS, St. Thomas's Hospital, London SE1 7EH,3 United Kingdom

Received 18 July 1997/Returned for modification 17 September 1997/Accepted 3 March 1998

Lactoferricin includes an 11-amino-acid amphipathic alpha-helical region which is exhibited on the outer surface of the amino-terminal lobe of lactoferrin. Synthetic peptides homologous to this region exhibited potent antibacterial activity against a selected range of both gram-negative and gram-positive bacteria. An analog synthesized with methionine substituted for proline at position 26, which is predicted to disrupt the helical region, abolished antibacterial activity against Escherichia coli and considerably reduced antibacterial activity against Staphylococcus aureus and an Acinetobacter strain. The mode of action of human lactoferrin peptide (HLP) 2 against E. coli serotype O111 (NCTC 8007) was established by using flow cytometry, surface plasmon resonance, and transmission electron microscopy. Flow cytometry was used to monitor membrane potential, membrane integrity, and metabolic processes by using the fluorescent probes bis-1,3-(dibutylbarbituric acid)-trimethine oxonol, propidium iodide, and carbonyl cyanide m-chlorophenylhydrazone, respectively. HLP 2 was found to act at the cell membrane, causing complete loss of membrane potential after 10 min and of membrane integrity within 30 min, with irreversible damage to the cell as shown by rapid loss of viability. The number of particles, measured by light scatter on the flow cytometer, dropped significantly, showing that bacterial lysis resulted. The peptide was shown to bind to E. coli O111 lipopolysaccharide by using surface plasmon resonance. Transmission electron microscopy revealed bacterial distortion, with the outer membrane becoming detached from the inner cytoplasmic membrane. We conclude that HLP 2 causes membrane disruption of the outer membrane, resulting in lysis, and that structural considerations are important for antibacterial activity.


* Corresponding author. Mailing address: Metalloprotein Research Group, Division of Biochemistry and Molecular Biology, UMDS, Guy's Hospital, London Bridge, London SE1 9RT, United Kingdom. Phone: (44) 171 955 4525. Fax: (44) 171 955 8881. E-mail: r.evans{at}umds.ac.uk.


Infect Immun, June 1998, p. 2434-2440, Vol. 66, No. 6
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.



This article has been cited by other articles:

  • Shalev, D. E., Rotem, S., Fish, A., Mor, A. (2006). Consequences of N-Acylation on Structure and Membrane Binding Properties of Dermaseptin Derivative K4-S4-(1-13). J. Biol. Chem. 281: 9432-9438 [Abstract] [Full Text]  
  • Hunter, H. N., Demcoe, A. R., Jenssen, H., Gutteberg, T. J., Vogel, H. J. (2005). Human Lactoferricin Is Partially Folded in Aqueous Solution and Is Better Stabilized in a Membrane Mimetic Solvent. Antimicrob. Agents Chemother. 49: 3387-3395 [Abstract] [Full Text]  
  • Radzishevsky, I. S., Rotem, S., Zaknoon, F., Gaidukov, L., Dagan, A., Mor, A. (2005). Effects of Acyl versus Aminoacyl Conjugation on the Properties of Antimicrobial Peptides. Antimicrob. Agents Chemother. 49: 2412-2420 [Abstract] [Full Text]  
  • Stokes, R. H., Oakhill, J. S., Joannou, C. L., Gorringe, A. R., Evans, R. W. (2005). Meningococcal Transferrin-Binding Proteins A and B Show Cooperation in Their Binding Kinetics for Human Transferrin. Infect. Immun. 73: 944-952 [Abstract] [Full Text]  
  • Shaper, M., Hollingshead, S. K., Benjamin, W. H. Jr., Briles, D. E. (2004). PspA Protects Streptococcus pneumoniae from Killing by Apolactoferrin, and Antibody to PspA Enhances Killing of Pneumococci by Apolactoferrin. Infect. Immun. 72: 5031-5040 [Abstract] [Full Text]  
  • Chapple, D. S., Hussain, R., Joannou, C. L., Hancock, R. E. W., Odell, E., Evans, R. W., Siligardi, G. (2004). Structure and Association of Human Lactoferrin Peptides with Escherichia coli Lipopolysaccharide. Antimicrob. Agents Chemother. 48: 2190-2198 [Abstract] [Full Text]  
  • Velliyagounder, K., Kaplan, J. B., Furgang, D., Legarda, D., Diamond, G., Parkin, R. E., Fine, D. H. (2003). One of Two Human Lactoferrin Variants Exhibits Increased Antibacterial and Transcriptional Activation Activities and Is Associated with Localized Juvenile Periodontitis. Infect. Immun. 71: 6141-6147 [Abstract] [Full Text]  
  • Majerle, A., Kidric, J., Jerala, R. (2003). Enhancement of antibacterial and lipopolysaccharide binding activities of a human lactoferrin peptide fragment by the addition of acyl chain. J Antimicrob Chemother 51: 1159-1165 [Abstract] [Full Text]  
  • Bengoechea, J. A., Brandenburg, K., Arraiza, M. D., Seydel, U., Skurnik, M., Moriyon, I. (2003). Pathogenic Yersinia enterocolitica Strains Increase the Outer Membrane Permeability in Response to Environmental Stimuli by Modulating Lipopolysaccharide Fluidity and Lipid A Structure. Infect. Immun. 71: 2014-2021 [Abstract] [Full Text]  
  • Navon-Venezia, S., Feder, R., Gaidukov, L., Carmeli, Y., Mor, A. (2002). Antibacterial Properties of Dermaseptin S4 Derivatives with In Vivo Activity. Antimicrob. Agents Chemother. 46: 689-694 [Abstract] [Full Text]  
  • Nibbering, P. H., Ravensbergen, E., Welling, M. M., van Berkel, L. A., van Berkel, P. H. C., Pauwels, E. K. J., Nuijens, J. H. (2001). Human Lactoferrin and Peptides Derived from Its N Terminus Are Highly Effective against Infections with Antibiotic-Resistant Bacteria. Infect. Immun. 69: 1469-1476 [Abstract] [Full Text]  
  • Lupetti, A., Paulusma-Annema, A., Welling, M. M., Senesi, S., van Dissel, J. T., Nibbering, P. H. (2000). Candidacidal Activities of Human Lactoferrin Peptides Derived from the N Terminus. Antimicrob. Agents Chemother. 44: 3257-3263 [Abstract] [Full Text]  
  • Levy, O. (2000). Antimicrobial proteins and peptides of blood: templates for novel antimicrobial agents. Blood 96: 2664-2672 [Abstract] [Full Text]  
  • Sallmann, F. R., Baveye-Descamps, S., Pattus, F., Salmon, V., Branza, N., Spik, G., Legrand, D. (1999). Porins OmpC and PhoE of Escherichia coli as Specific Cell-surface Targets of Human Lactoferrin. BINDING CHARACTERISTICS AND BIOLOGICAL EFFECTS. J. Biol. Chem. 274: 16107-16114 [Abstract] [Full Text]  
  • Hancock, R. E. W., Chapple, D. S. (1999). Peptide Antibiotics. Antimicrob. Agents Chemother. 43: 1317-1323 [Full Text]  
  • Wakabayashi, H., Matsumoto, H., Hashimoto, K., Teraguchi, S., Takase, M., Hayasawa, H. (1999). N-Acylated and D Enantiomer Derivatives of a Nonamer Core Peptide of Lactoferricin B Showing Improved Antimicrobial Activity. Antimicrob. Agents Chemother. 43: 1267-1269 [Abstract] [Full Text]