title: Differential binding of L- vs. D-isomers of cationic antimicrobial peptides to the biofilm exopolysaccharide alginate.
abstract: Alginate is a biofilm exopolysaccharide secreted by the opportunistic pathogen Pseudomonas aeruginosa that acts to prevent the diffusion of antibiotics toward the bacterial cell membrane. Cationic antimicrobial peptides (CAPs) have been increasingly recognized as a viable alternative for prospective antimicrobial agents. The D-isomer chiral counterparts of active L-isomer CAPs tend to show slightly greater antimicrobial activities because bacteria lack proteases to hydrolyze the unnatural D-isomers. Using an enantiomeric pair of synthetic CAPs designed in our laboratory (L-4Leu in the sequence KKKKKKALFALWLAFLA-NH2 and its D-analog D-4Leu), we studied the binding and interactions of Lvs. D-isomers of CAPs with alginate using circular dichroism and Raman spectroscopic techniques. We found that the peptide D-4Leu underwent a more rapid structural transition over time from an initial alginate-induced α-helical conformation to a less soluble β-sheet conformation than L-4Leu, indicating that the D-isomer of this peptide has a relatively greater affinity for alginate. Through Raman spectroscopy it was observed that Raman modes at 1297 cm-1 and 1453 cm-1 wavenumbers were found to differ between the spectra obtained from the insoluble complexes formed between L-4Leu vs. D-4Leu and alginate. These modes were tentatively assigned to CH, and CH3 deformation modes, respectively. Our findings reveal previously undetected subtleties in the binding of this diastereomeric pair of peptides in the microenvironment of a biofilm exopolysaccharide, and provide guidelines for future development of antimicrobial peptides.
authors: Division of Molecular Structure & Function, Research Institute, Hospital for Sick Children, Toronto M5G 1X8.
authors: Lois M
authors: LM
authors: Yin
authors: Soyoung
authors: S
authors: Lee
authors: Jacky S W
authors: JS
authors: Mak
authors: Amr S
authors: AS
authors: Helmy
authors: Charles M
authors: CM
authors: Deber
mesh_terms: D000464:Alginates / Q000378:metabolism*; D000595:Amino Acid Sequence; D000890:Anti-Infective Agents / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D018441:Biofilms / Q000187:drug effects; D002942:Circular Dichroism; D020723:Glucuronic Acid / Q000378:metabolism; D006603:Hexuronic Acids / Q000378:metabolism; D008969:Molecular Sequence Data; D011135:Polysaccharides, Bacterial / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D011550:Pseudomonas aeruginosa / Q000187:drug effects; D013059:Spectrum Analysis, Raman; D013237:Stereoisomerism
chemical_list: D000464:Alginates; D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D006603:Hexuronic Acids; D011135:Polysaccharides, Bacterial; C064841:exopolysaccharide, Pseudomonas; D020723:Glucuronic Acid