Peptide-membrane interactions studied by a new phospholipid/polydiacetylene colorimetric vesicle assay.
Interactions between peptides and lipid membranes play major roles in numerous physiological processes, such as signaling, cytolysis, formation of ion channels, and cellular recognition. We describe a new colorimetric technique for studying peptide-membrane interactions. The new assay is based on supramolecular assemblies composed of phospholipids embedded in a matrix of polydiacetylene (PDA) molecules. The phospholipid/PDA vesicle solutions undergo visible color changes upon binding of membrane peptides. Experiments utilizing various analytical techniques confirm that the blue-to-red color transitions of the phospholipid/PDA vesicles are directly related to adoption of helical conformations by the peptides and their association with the lipids. Spectroscopic data indicate that the colorimetric transitions are correlated with important molecular parameters, such as the degree of penetration of the peptides into lipid bilayers, and the mechanisms of peptide-lipid binding. The results suggest that the new colorimetric assay could be utilized for studying interactions and organization of membrane peptides.
Kolusheva
S
S
Department of Chemistry and Stadler Minerva Center for Mesoscopic, Macromolecular Engineering, Ben Gurion University of the Negev, Beersheva 84105, Israel.
Shahal
T
T
Jelinek
R
R
D000114:Acetylene / Q000031:analogs & derivatives* / Q000737:chemistry*; D000408:Alamethicin / Q000737:chemistry; D000595:Amino Acid Sequence; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry; D002942:Circular Dichroism; D003124:Colorimetry / Q000379:methods*; D004134:Dimyristoylphosphatidylcholine / Q000737:chemistry; D004578:Electron Spin Resonance Spectroscopy; D008081:Liposomes / Q000737:chemistry*; D008555:Melitten / Q000737:chemistry; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D010455:Peptides / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry*; D000078789:Polyacetylene Polymer; D011108:Polymers / Q000737:chemistry*; D053279:Polyynes; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D023181:Antimicrobial Cationic Peptides; D008081:Liposomes; D010455:Peptides; D010743:Phospholipids; D000078789:Polyacetylene Polymer; D011108:Polymers; D008555:Melitten; D053279:Polyynes; D000408:Alamethicin; C082361:polydiacetylene; D000114:Acetylene; D004134:Dimyristoylphosphatidylcholine