This article describes efforts to build a model biological membrane at a surface of a gold electrode. In this architecture, the membrane may be exposed to static electric fields on the order of 107 to 108 V m−1. These fields are comparable in magnitude to the static electric field acting on a natural biological membrane. The field may be conveniently used to manipulate organic molecules within the membrane. By turning a knob on the control instrument one can deposit or lift the membrane from the gold surface. Electrochemical techniques can be used to control the physical state of the film while the infrared reflection absorption spectroscopy (IRRAS), surface imaging by STM and AFM and neutron scattering techniques can be employed to study conformational changes of organic molecules and their ordering within the membrane. This is shown on examples of membranes built of a simple zwitterionic phospholipid such as 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) and a mixed membrane composed of DMPC and cholesterol. The results illustrate the tremendous effect of cholesterol on the membrane structure. Two methods of membrane deposition at the electrode surface, namely by unilamellar vesicles fusion and using the Langmuir–Blodgett technique, are compared. Applications of these model systems to study interactions of small antibiotic peptides with lipids are discussed.
10 December 2012 in Proceedings of the National Academy of Sciences
We present direct visualization of pores formed by alamethicin (Alm) in a matrix of phospholipids using electrochemical scanning tunneling microscopy (EC-STM). High-resolution EC-STM images show individual peptide molecules forming channels. The channels are not dispersed randomly in the monolayer but agglomerate forming 2D nanocrystals with a hexagonal lattice in which the average channel–channel distance is 1.90 ± 0.1 nm. The STM images suggest that each Alm is shared between the two adjacent channels. Every channel consists of six Alm molecules. Three or four of these molecules have the hydrophilic group oriented toward the center of the channel allowing for water column formation inside the channel. The dimensions of the central pore in the images are consistent with the dimension of the water column in a model of hexameric pore proposed in the literature. The images obtained in this work validate the barrel-stave model of the pore formed in phospholipid membranes by amphiphatic peptides. They also provide direct evidence for cluster formation by such pores.