title: Alginate as an auxiliary bacterial membrane: binding of membrane-active peptides by polysaccharides.
abstract: The chronicity of Pseudomonas aeruginosa infections in cystic fibrosis (CF) patients is characterized by overproduction of the exopolysaccharide alginate, in which biofilm bacteria are embedded. Alginate apparently contributes to the antibiotic resistance of bacteria in this form by acting as a diffusion barrier to positively charged antimicrobial agents. We have been investigating cationic antimicrobial peptides (CAPs) (prototypic sequence: KKAAAXAAAAAXAAWAAXAAAKKKK-NH(2), where X is any of the 20 commonly occurring amino acids) that were originally designed as transmembrane mimetic peptides. Peptides of this group above a specific hydrophobicity threshold insert spontaneously into membranes and have antibacterial activity at micromolar concentrations. While investigating the molecular basis of biofilm resistance to peptides, we found that the anionic alginate polysaccharide induces conformational changes in the most hydrophobic of these peptides typically associated with insertion of such peptides into membrane environments [Chan et al., J. Biol. Chem. (2004) vol. 279, pp. 38749-38754]. Through a combination of experiments measuring release of the fluorescent dye calcein from phospholipid vesicles, peptide interactions with vesicles in the presence and absence of alginate, and affinity of peptides for alginate as a function of net peptide core hydrophobicity, we show here that alginate offers a microenvironment that provides a protective mechanism for the encased bacteria by both binding and promoting the self-association of the CAPs. The overall results indicate that hydrophilic alginate polymers contain a significant hydrophobic compartment, and behave as an 'auxiliary membrane' for bacteria, thus identifying a unique protective role for biofilm exopolysaccharide matrices.
authors.affiliation: Division of Structural Biology and Biochemistry, Research Institute, The Hospital for Sick Children, Toronto, Ontario, Canada.
authors.forename: C
authors.initials: C
authors.lastname: Chan
authors.forename: L L
authors.initials: LL
authors.lastname: Burrows
authors.forename: C M
authors.initials: CM
authors.lastname: Deber
mesh_terms: D000464:Alginates / Q000737:chemistry*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D018441:Biofilms; D004058:Diffusion; D005452:Fluoresceins / Q000737:chemistry; D020723:Glucuronic Acid / Q000737:chemistry*; D006603:Hexuronic Acids / Q000737:chemistry*; D008081:Liposomes / Q000737:chemistry; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D011485:Protein Binding
chemical_list: D000464:Alginates; D023181:Antimicrobial Cationic Peptides; D005452:Fluoresceins; D006603:Hexuronic Acids; D008081:Liposomes; D011135:Polysaccharides, Bacterial; C064841:exopolysaccharide, Pseudomonas; D020723:Glucuronic Acid; C007740:fluorexon