Surface supported bilayer platform for studies of lateral association of proteins in membranes (Mini Review)

Abstract

Here, the authors review how surface supported bilayers can be engineered and how Förster resonance energy transfer (FRET) can be used to quantify interactions between transmembrane peptides in these bilayers. The requirements for the surface supported platform are (1) lateral mobility of the peptides, (2) transmembrane orientation of the peptides, and (3) capabilities for FRET measurements. To satisfy these requirements, a new assembly method, termed “directed assembly” was developed. This assembly method could have broad utility in basic studies of proteins in membranes and in biotechnological applications.

Fluorescent Analogs of Biomolecular Building Blocks: Design, Properties, and Applications

“…FRET has also been used to elucidate folding and dynamics of RNA,25 as well as the sequence dependent structure, stability and dynamics of nucleosomes 26. Membrane researchers have used FRET to study, for example, microdomain formation19 and trans-membrane peptides in surface supported bilayers 27…”

Effect of a Polymer Cushion on the Electrical Properties and Stability of Surface-Supported Lipid Bilayers

“…Biomimetic membranes on planar solid supports are being developed to study the structure, function, and stability of membrane proteins in a controlled manner. − The assembly of such membranes on conductive substrates allows electrical detection of ion permeation by electrochemical impedance spectroscopy (EIS). − The architecture of supported membranes often includes a polymeric cushion, usually poly(ethylene glycol) (PEG), to raise the bilayer above the substrate. , It is generally believed that this polymeric cushion facilitates the incorporation of membrane proteins with cytoplasmic domains by preventing denaturing contacts between the protein and the substrate. , However, a systematic characterization and optimization of the PEG cushions in such studies is rarely performed.…”

Fluorophores, environments, and quantification techniques in the analysis of transmembrane helix interaction using FRET

“…For example, Engelman and coworkers and Schneider and coworkers have used FRET to study the dimerization of the model system GpA and its sequence variants in different membrane mimetic environments. Hristova and coworkers have contributed significantly to the field of FRET in membrane proteins by developing new experimental methodologies, testing different bilayer environments, and improving on existing formalisms for FRET calculations to investigate the dimerization of proteins of the receptor tyrosine kinase family . Deber and coworkers have studied oligomerization of natural and model peptides to understand the structure–function relationship of receptors, ion pumps, and other biological systems and to investigate the rules that govern membrane protein folding .…”

“…The most widely used membrane mimetics for studying membrane protein oligomerization are lipid bilayers and detergent micelles. Bilayers, which are a more native‐like environment, can be synthetically prepared by different methods: in the form of supported bilayers, multilamellar vesicles (MLVs), and giant (∼500 nm), large unilamellar vesicles (LUV; ∼100 nm), or small unilamellar vesicles (SUV; ∼30 to 50 nm). As discussed later, these alternative forms may be best suited to different experimental requirements .…”