Microfluidic-Derived Detection of Protein-Facilitated Copper Flux Across Lipid Membranes

Abstract

Measurement of protein-facilitated copper flux across biological membranes is a considerable challenge. Here, we demonstrate a straightforward microfluidic-derived approach for visualization and measurement of membranous Cu flux. Giant unilamellar vesicles, reconstituted with the membrane protein of interest, are prepared, surface-immobilized, and assessed using a novel quencher–sensor reporter system for detection of copper. With the aid of a syringe pump, the external buffer is exchanged, enabling consistent and precise exchange of solutes, without causing vesicle rupture or uneven local metal concentrations brought about by rapid mixing. This approach bypasses common issues encountered when studying heavy metal-ion flux, thereby providing a new platform for in vitro studies of metal homeostasis aspects that are critical for all cells, health, and disease.

Lipid vesicle-based molecular robots

“… 298 On the other hand, molecular robots with sensors and computers may be able to fill this gap. Previous studies have reported that GUVs with nanopores are useful for the detection of K + , 56 Cu 2+ , 299 and Cl − . 300 Recently, a unique strategy using GUVs with a DNA nanopore and circuit offered by Peng et al suggested switchable detection of Sr 2+ (ref.…”