Abstract
Antibacterial peptides are part of the innate immune system in a variety of different species including humans. Some of these peptides have also been shown to have effects on immune competent cells such as professional phagocytes. We have recently shown that a cecropin-like peptide from Helicobacter pylori, Hp(2–20), in addition to being bactericidal possesses proinflammatory effects and can recruit and activate neutrophils as well as monocytes. It is well established that cecropins have the ability to adopt amphipathic α-helices, which is thought to be required for their bactericidal activity. In this study we show the same structural requirements for Hp(2–20). Breaking the helical structure of Hp(2–20) reduced the antibacterial effect and abolished its proinflammatory activity. A C-terminal truncated cecropin A peptide that highly resembles Hp(2–20) failed to activate neutrophils and computer-based structural simulations revealed a difference between the two peptides in the stability of their helical structures. A hybrid peptide with amino acid substitutions stabilizing the α-helical structure of the truncated cecropin A peptide did not introduce any proinflammatory activity; the bactericidal activity was, however, increased. We thus conclude that the proinflammatory effect of Hp(2–20) is a unique sequence-specific feature of the peptide and the ability to adopt a stable amphipathic helix is a necessary but not sufficient criterion for the functional dualism of the peptide.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Boman, H. G. 2000. Innate immunity and the normal microflora. Immunol Rev. 173:5–16.
Epand, R. M. and H. J. Vogel. 1999. Diversity of antimicrobial peptides and their mechanisms of action. Biochim. Biophys. Acta. 1462:11–28.
Putsep, K., C. I. Branden, H. G. Boman, and S. Normark. 1999. Antibacterial peptide from H. pylori. Nature. 398:671–672.
Allen, L. A. 2000. Modulating phagocyte activation: the pros and cons of Helicobacter pylori virulence factors. J. Exp. Med. 191:1451–1454.
Gudmundsson, G. H., and B. Agerberth. 1999. Neutrophil antibacterial peptides, multifunctional effector molecules in the mammalian immune system. J. Immunol. Methods 232:45–54.
Graham, D. Y. 2000. Helicobacter pylori infection is the primary cause of gastric cancer. J Gastroenterol. 35:90–97.
Betten, A., J. Bylund, T. Cristophe, F. Boulay, A. Romero, K. Hellstrand, and C. Dahlgren, C. 2001. A proinflammatory peptide from Helicobacter pylori activates monocytes to induce lymphocyte dysfunction and apoptosis. J. Clin. Invest. 108:1221–1228.
Bylund, J., T. Christophe, F. Boulay, T. Nystrom, A. Karlsson, and C. Dahlgren. 2001. Proinflammatory activity of a cecropin-like antibacterial peptide from Helicobacter pylori. Antimicrob. Agents Chemother. 45:1700–1704.
de Paulis, A., N. Prevete, I. Fiorentino, A. F. Walls, M. Curto, A. Petraroli, V. Castaldo, P. Ceppa, R. Fiocca, and G. Marone. 2004. Basophils infiltrate human gastric mucosa at sites of Helicobacter pylori infection, and exhibit chemotaxis in response to H. pylori-derived peptide Hp(2–20). J. Immunol. 172:7734–7743.
Bylund, J. 2002. Recognition by leukocyte formyl peptide receptors: promiscuous binding or pattern recognition? Medical dissertation. Göteborg University, Göteborg.
Bylund, J., A. Karlsson, F. Boulay, and C. Dahlgren, 2002. Lipopolysaccharide-induced granule mobilization and priming of the neutrophil response to Helicobacter pylori peptide Hp(2–20), which activates formyl peptide receptor-like 1. Infect. Immun. 70:2908–2914.
Boyum, A., D. Lovhaug, L. Tresland, and E. M. Nordlie, 1991. Separation of leucocytes: Improved cell purity by fine adjustments of gradient medium density and osmolality. Scand. J. Immunol. 34:697–712.
Dahlgren, C., and A. Karlsson, 1999. Respiratory burst in human neutrophils. J. Immunol. Methods 232:3–14.
Hultmark, D., A. Engstrom, K. Andersson, H. Steiner, H. Bennich, and H. G. Boman, 1983. Insect immunity. Attacins, a family of antibacterial proteins from Hyalophora cecropia. Embo. J. 2:571–576.
Andreu, D., R. B. Merrifield, H. Steiner, and H. G. Boman, 1985. N-terminal analogues of cecropin A: Synthesis, antibacterial activity, and conformational properties. Biochemistry 24:1683–1688.
Clark, R. A. 1999. Activation of the neutrophil respiratory burst oxidase. J. Infect. Dis. 179(Suppl 2):S309–S317.
Holak, T. A., A. Engstrom, P. J. Kraulis, G. Lindeberg, H. Bennich, T. A. Jones, A. M. Gronenborn, and G. M. Clore. 1988. The solution conformation of the antibacterial peptide cecropin A: A nuclear magnetic resonance and dynamical simulated annealing study. Biochemistry 27:7620–1729.
Shai, Y. 1999. Mechanism of the binding, insertion and destabilization of phospholipid bilayer membranes by alpha-helical antimicrobial and cell non-selective membrane-lytic peptides. Biochim. Biophys. Acta 1462:55–70.
Christensen, B., J. Fink, R. B. Merrifield, and D. Mauzerall. 1988. Channel-forming properties of cecropins and related model compounds incorporated into planar lipid membranes. Proc. Natl. Acad. Sci. USA 85:5072–5076.
Gazit, E., I. R. Miller, P. C. Biggin, M. S. Sansom, and Y. Shai. 1996. Structure and orientation of the mammalian antibacterial peptide cecropin P1 within phospholipid membranes. J. Mol. Biol. 258:860–870.
De Lucca, A. J., T. J. Jacks, and K. A. Brogden. 1995. Binding between lipopolysaccharide and cecropin A. Mol. Cell Biochem. 151:141–148.
Ye, R. D. and F. Boulay. 1997. Structure and function of leukocyte chemoattractant receptors. Adv. Pharmacol. 39:221–289.
Fu, H., L. Bjorkman, P. Janmey, A. Karlsson, J. Karlsson, C. Movitz, and C. Dahlgren. 2004. The two neutrophil members of the formylpeptide receptor family activate the NADPH-oxidase through signals that differ in sensitivity to a gelsolin derived phosphoinositide-binding peptide. BMC Cell Biol. 5:50.
Fu, H., J. Bylund, A. Karlsson, S. Pellme, and C. Dahlgren. 2004. The mechanism for activation of the neutrophil NADPH-oxidase by the peptides formyl-Met-Leu-Phe and Trp-Lys-Tyr-Met-Val-Met differs from that for interleukin-8. Immunology 112:201–210.
Dahlgren, C., T. Christophe, F. Boulay, P. N. Madianos, M. J. Rabiet, and A. Karlsson. 2000. The synthetic chemoattractant Trp-Lys-Tyr-Met-Val-DMet activates neutrophils preferentially through the lipoxin A(4) receptor. Blood 95:1810–1818.
Le, Y., W. Gong, H. L. Tiffany, A. Tumanov, S. Nedospasov, W. Shen, N. M. Dunlop, J. L. Gao, P. M. Murphy, J. J. Oppenheim, and J. M. Wang. 2001. Amyloid (beta)42 activates a G-protein-coupled chemoattractant receptor, FPR-like-1. J. Neurosci. 21:RC123.
Hartt, J. K., T. Liang, A. Sahagun-Ruiz, J. M. Wang, J. L. Gao, and P. M. Murphy. 2000. The HIV-1 cell entry inhibitor T-20 potently chemoattracts neutrophils by specifically activating the N-formylpeptide receptor. Biochem. Biophys. Res. Commun. 272:699–704.
Deng, X., H. Ueda, S. B. Su, W. Gong, N. M. Dunlop, J. L. Gao, P. M. Murphy, and J. M. Wang. 1999. A synthetic peptide derived from human immunodeficiency virus type 1 gp 120 downregulates the expression and function of chemokine receptors CCR5 and CXCR4 in monocytes by activating the 7-transmembrane G-protein-coupled receptor FPRL1/LXA4R. Blood 94:1165–1173.
Chiang, N., I. M. Fierro, K. Gronert, and C. N. Serhan. 2000. Activation of lipoxin A(4) receptors by aspirin-triggered lipoxins and select peptides evokes ligand-specific responses in inflammation. J. Exp. Med. 191:1197–1208.
Badolato, R., J. M. Wang, W. J. Murphy, A. R. Lloyd, D. F. Michiel, L.L Bausserman, D. J. Kelvin, and J. J. Oppenheim. 1994. Serum amyloid A is a chemoattractant: Induction of migration, adhesion, and tissue infiltration of monocytes and polymorphonuclear leukocytes. J. Exp. Med. 180:203–209.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Fu, H., Björstad, Å., Dahlgren, C. et al. A Bactericidal Cecropin-A Peptide with a Stabilized α-Helical Structure Possess an Increased Killing Capacity But No Proinflammatory Activity. Inflammation 28, 337–343 (2004). https://doi.org/10.1007/s10753-004-6644-9
Issue Date:
DOI: https://doi.org/10.1007/s10753-004-6644-9