title: Thermodynamic profiling of peptide membrane interactions by isothermal titration calorimetry: a search for pores and micelles.
abstract: Antimicrobial peptides are known to interact strongly with negatively charged lipid membranes, initially by peripheral insertion of the peptide into the bilayer, which for some antimicrobial peptides will be followed by pore formation, and successive solubilization of the membranes resulting in mixed peptide-lipid micelles. We have investigated the mode of action of the antimicrobial peptide mastoparan-X using isothermal titration calorimetry (ITC) and cryo-transmission electron microscopy (cryo-TEM). The results show that mastoparan-X induces a range of structural transitions of POPC/POPG (3:1) lipid membranes at different peptide/lipid ratios. It has been established that ITC can be used as a fast method for localizing membrane transitions and when combined with DLS and cryo-TEM can elucidate structural changes, including the threshold for pore formation and micellation. Cryo-TEM was employed to confirm the structural changes associated with the thermodynamic transitions found by ITC. The pore-formation process has furthermore been investigated in detail and the thermodynamic parameters of pore formation have been characterized using a system-specific temperature where the enthalpy of peptide partitioning becomes zero (T(zero)). This allows for an exclusive study of the pore-formation process. The use of ITC to find T(zero) allows for characterization of the thermodynamic parameters of secondary processes on lipid membranes.
authors: Department of Micro- and Nanotechnology, Technical University of Denmark, Lyngby, Denmark.
authors: J R
authors: JR
authors: Henriksen
authors: T L
authors: TL
authors: Andresen
mesh_terms: D002151:Calorimetry / Q000379:methods*; D020285:Cryoelectron Microscopy; D036341:Intercellular Signaling Peptides and Proteins; D008567:Membranes, Artificial; D008823:Micelles; D010455:Peptides / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry; D010715:Phosphatidylglycerols / Q000737:chemistry; D016062:Porosity; D013816:Thermodynamics; D017186:Titrimetry
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D008567:Membranes, Artificial; D008823:Micelles; D010455:Peptides; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; C071123:mastoparan X; C060037:1-palmitoyl-2-oleoylglycero-3-phosphoglycerol; C028694:1-palmitoyl-2-oleoylphosphatidylcholine