Arrayed lipid bilayer chambers allow single-molecule analysis of membrane transporter activity

Area of Science:

• Biophysics
• Biochemistry
• Nanotechnology

Background:

• Femtolitre chamber arrays are valuable for sensitive biological assays.
• Current femtolitre chamber arrays are limited to aqueous reactions.
• Analyzing membrane transport requires specialized platforms.

Purpose of the Study:

• To develop a novel platform for single-molecule analysis of membrane transport.
• To expand the capabilities of femtolitre chamber arrays beyond aqueous solutions.
• To investigate passive and active transport mechanisms of membrane proteins.

Main Methods:

• Development of an arrayed lipid bilayer chamber system (ALBiC) with sub-million femtolitre chambers.
• Each chamber is sealed with a stable 4-μm-diameter lipid bilayer membrane.
• Reconstitution of chambers with membrane transporter proteins (α-hemolysin, F0F1-ATP synthase).

Main Results:

• ALBiC chambers exhibit stochastic and quantized transporting activities.
• Demonstrated single-molecule analysis of passive and active membrane transport.
• Successful reconstitution and functional observation of membrane transporter proteins.

Conclusions:

• The ALBiC system enables single-molecule analysis of membrane transport.
• This platform broadens the versatility of femtolitre chamber arrays.
• ALBiC facilitates novel applications for understanding membrane protein function.