Binding orientation and activity determinants of the antimicrobial peptide cryptdin-4 revealed by potency of mutants.
Cryptdin-4 is a beta-sheet antimicrobial peptide of the defensin family that is found in the immune system of mice. Several structure-activity studies of this peptide have previously been conducted, but none have been based on residue-membrane interactions as part of an overall hypothesis on the peptide's orientation in the membrane. We pursue this valuable approach by first using previously reported NMR structural data to propose a membrane-bound orientation of the peptide. Four mutants are then strategically designed to modulate membrane perturbative activity in a manner consistent with the proposed binding orientation. Membrane perturbation is evaluated using a simple fluorescence-based vesicle leakage assay using POPG to form the model membrane. Effects of peptide mutations are found to be consistent with the suggested binding orientation. This approach is successfully used to create synthetic peptides with enhanced or diminished ability to perturb membranes and also yields insights on the nature of peptide-membrane interactions.
Cummings
Jason E
JE
Department of Chemical Engineering, Princeton University, Princeton, NJ 08544, Unites States.
Vanderlick
T Kyle
TK
D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D001665:Binding Sites; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D008567:Membranes, Artificial; D051379:Mice; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D010715:Phosphatidylglycerols / Q000737:chemistry; D017433:Protein Structure, Secondary; D013499:Surface Properties; D023084:alpha-Defensins / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics
D023181:Antimicrobial Cationic Peptides; D008567:Membranes, Artificial; D010715:Phosphatidylglycerols; D023084:alpha-Defensins; C401847:cryptdin 4, mouse; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol