title: Lipopolysaccharide structure determines ionic and hydrophobic binding of a cationic antimicrobial neutrophil granule protein.
abstract: Bactericidal activity and binding of a 57,000-dalton cationic antimicrobial neutrophil granule protein (CAP57) are determined by the presence on bacteria of O-antigen polysaccharide chains and the availability of negatively charged groups in the lipid A region, the inner core region, or both regions of lipopolysaccharide. Polymyxin B (PMB)-resistant mutants with well-defined alterations in lipid A structure and charge (pmrA) are also more resistant to CAP57. We used biologically active radioiodinated CAP57 to study the characteristics and kinetics of binding to a sensitive Rb lipopolysaccharide chemotype, Salmonella typhimurium SH9178, and the relatively resistant pmrA mutant strain SH7426. Binding occurred rapidly and was specific and saturable. Because CAP57 appears to be bound in a manner similar to that of PMB, competition binding studies were performed. Excess PMB did compete with CAP57 for binding to SH9178. Nonapeptide, a polycationic derivative of PMB that has lost its hydrophobic portions, demonstrated a marked decrease in ability to compete for binding with CAP57 compared with PMB. This demonstrated the importance of hydrophobic binding in the interaction of CAP57 with the microbial surface. Thus, we have shown that binding of CAP57 to SH9178 is specific, saturable, and similar to binding of PMB. Both hydrophobic and ionic properties of CAP57 appear to be necessary for binding.
authors.affiliation: Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia 30322.
authors.forename: M M
authors.initials: MM
authors.lastname: Farley
authors.forename: W M
authors.initials: WM
authors.lastname: Shafer
authors.forename: J K
authors.initials: JK
authors.lastname: Spitznagel
mesh_terms: D023181:Antimicrobial Cationic Peptides; D001665:Binding Sites / Q000187:drug effects; D001667:Binding, Competitive; D001770:Blood Bactericidal Activity / Q000187:drug effects; D001798:Blood Proteins / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides / Q000097:blood; D009504:Neutrophils / Q000378:metabolism* / Q000382:microbiology; D009842:Oligopeptides / Q000494:pharmacology; D011112:Polymyxin B / Q000494:pharmacology; D012486:Salmonella typhimurium / Q000378:metabolism*; D013329:Structure-Activity Relationship
chemical_list: D023181:Antimicrobial Cationic Peptides; D001798:Blood Proteins; D008070:Lipopolysaccharides; D009842:Oligopeptides; C053368:cationic antimicrobial protein 57, human; D011112:Polymyxin B