New Biophysical Methods to Study the Membrane Activity of Bcl-2 Proteins

Abstract

The proteins of Bcl-2 family are key regulators of apoptosis. Many Bcl-2 proteins have the unique ability to switch between two possible conformations, soluble in the cytosol or associated to cellular membranes. Importantly, their membrane-inserted form is the main responsible for their apoptotic function. Unfortunately, there are only a limited number of methods available to study the membrane activity of these proteins. Here, we present a methodology to study protein binding to membranes and membrane permeabilization at the single vesicle level. It is based on purified proteins and giant unilamellar vesicles and involves directly visualization of the process with a confocal microscope. This approach allows for the characterization of the membrane activity of the Bcl-2 proteins (or of any other pore-forming molecule) with unprecedented detail.