title: Helix induction in antimicrobial peptides by alginate in biofilms.
abstract: Bacterial exopolysaccharides provide protection against phagocytosis, opsonization, and dehydration and act as a major structural component of the extracellular matrix in biofilms. They contribute to biofilm-related resistance by acting as a diffusion barrier to positively charged antimicrobial agents including cationic antimicrobial peptides (CAPs). We previously created novel CAPs consisting of a nonamphipathic hydrophobic core flanked by Lys residues and containing a Trp residue in the hydrophobic segment as a fluorescent probe. Peptides of this type above a specific hydrophobicity threshold insert spontaneously into membranes and have antimicrobial activity against Gram-positive and Gram-negative bacteria at micromolar concentrations. Here we show that alginate, a polymer of beta-d-mannuronate and alpha-l-guluronate secreted by the cystic fibrosis pathogen Pseudomonas aeruginosa, induces an alpha-helical conformation detected by circular dichroism spectroscopy and blue shifts in Trp fluorescence maxima in peptides above the hydrophobicity threshold, changes typically observed upon association of such peptides with nonpolar (membrane) environments. Parallel effects were observed in the archetypical CAPs magainin II amide and cecropin P1. Fluorescence resonance energy transfer studies indicated that alginate induces peptide-peptide association only in peptides above the hydrophobicity threshold, suggesting that the hydrophilic alginate polymer behaves as an "auxiliary membrane" for the bacteria, demonstrating a unique protective role for biofilm matrices against CAPs.
authors.affiliation: Division of Structural Biology and Biochemistry, Research Institute, The Hospital for Sick Children, Toronto, Ontario M5G 1X8 Canada.
authors.forename: Celine
authors.initials: C
authors.lastname: Chan
authors.forename: Lori L
authors.initials: LL
authors.lastname: Burrows
authors.forename: Charles M
authors.initials: CM
authors.lastname: Deber
mesh_terms: D000464:Alginates / Q000737:chemistry*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry / Q000494:pharmacology*; D018441:Biofilms; D002942:Circular Dichroism; D004058:Diffusion; D031541:Fluorescence Resonance Energy Transfer; D006603:Hexuronic Acids / Q000737:chemistry; D008239:Lysine / Q000737:chemistry; D018613:Microscopy, Confocal; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry; D011108:Polymers / Q000737:chemistry; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011550:Pseudomonas aeruginosa / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D014364:Tryptophan / Q000737:chemistry
chemical_list: D000464:Alginates; D023181:Antimicrobial Cationic Peptides; D006603:Hexuronic Acids; D010455:Peptides; D011108:Polymers; C007896:guluronic acid; D014364:Tryptophan; C008324:mannuronic acid; D008239:Lysine; C070016:cecropin P1-LI