Lactobionamide-based fluorinated detergent for functional and structural stabilization of membrane proteins

Area of Science:

• Biochemistry
• Structural Biology
• Drug Discovery

Background:

• Membrane proteins (MPs) are vital drug targets, but their stabilization and solubilization present challenges.
• Conventional detergents show variable success in maintaining MP functionality and stability.
• Fluorinated surfactants (FSs) show promise for MP stabilization, yet their solubilization capabilities are less explored.

Purpose of the Study:

• To introduce and evaluate FLAC6, a novel fluorinated lactobionamide-based detergent, for solubilizing and stabilizing membrane proteins.
• To assess the impact of FLAC6 on the purity, homogeneity, and functionality of solubilized MPs.
• To investigate the overstabilization effects of FLAC6 on membrane proteins.

Main Methods:

• FLAC6 was used to solubilize native adenosine receptor A2A, AcrB, and BmrA.
• Purified MPs were analyzed for purity, homogeneity, and aggregation.
• MP functionality was assessed post-solubilization.
• Thermal denaturation (Tm) was measured to evaluate protein stabilization.

Main Results:

• FLAC6 efficiently solubilized three distinct membrane proteins: A2A, AcrB, and BmrA.
• Affinity-purified MPs were highly pure, homogenous, and free of aggregates.
• MP functionality was well-preserved.
• FLAC6 significantly increased the Tm of A2A, AcrB, and BmrA by 7°C, ~9°C, and ~23°C, respectively, compared to a reference detergent.

Conclusions:

• FLAC6 is an effective detergent for the functional and structural stabilization of diverse membrane proteins.
• This detergent offers a new strategy for stabilizing challenging MPs, enhancing their druggability for unmet medical needs.
• FLAC6 provides a valuable tool for advancing research in membrane protein structure, function, and drug discovery.