Transport assays for sterol-binding proteins: Stopped-flow fluorescence methods for investigating intracellular cholesterol transport mechanisms of NPC2 protein

Abstract

In this chapter we describe the use of stopped flow fluorescence spectroscopy to analyze the kinetic mechanisms of protein mediated cholesterol transfer to, from, and between model membranes. These assays allow for the detection of protein–membrane interactions that may occur during cholesterol transfer by simply modifying donor or acceptor concentrations, membrane composition, or buffer properties, and analyzing resultant transfer rates.