Cholesterol Chip for the Study of Cholesterol-Protein Interactions Using SPR

Cholesterol, an important lipid in animal membranes, binds to hydrophobic pockets within many soluble proteins, transport proteins and membrane bound proteins. The study of cholesterol-protein interactions in aqueous solutions is complicated by cholesterol's low solubility and often requires organic co-solvents or surfactant additives. We report the synthesis of a biotinylated cholesterol and immobilization of this derivative on a streptavidin chip. Surface plasmon resonance (SPR) was then used to measure the kinetics of cholesterol interaction with cholesterol-binding proteins, hedgehog protein and tyrosine phosphatase 1B.

Automated summary

Cholesterol, an important lipid in animal cell membranes, can bind to various soluble proteins, transport proteins, and membrane-bound proteins. However, studying the interactions between cholesterol and proteins in aqueous solutions is challenging due to the low solubility of cholesterol, often necessitating the use of organic co-solvents or surfactant additives. In this study, a biotinylated cholesterol derivative was synthesized and immobilized on a streptavidin chip for analysis using surface plasmon resonance (SPR). The kinetics of cholesterol interactions with cholesterol-binding proteins, hedgehog protein and tyrosine phosphatase 1B, were determined.