Assessing water permeability of aquaporins in a proteoliposome-based stopped-flow setup

• Aquaporins (AQPs) are water channels embedded in the cell membrane that are critical in maintaining water homeostasis. We describe a protocol for determining the water permeation capacity of AQPs reconstituted into proteoliposomes. Using a stopped-flow setup, AQP embedded in proteoliposomes are exposed to an osmogenic gradient that triggers water flux. The consequent effects on proteoliposome size can be tracked using the fluorescence of an internalized fluorophore. This enables controlled characterization of water flux by AQPs. For complete details on the use and execution of this protocol, please refer to Kitchen et al. (2020).

Subject headings

TEKNIK OCH TEKNOLOGIER  -- Miljöbioteknik -- Vattenbehandling (hsv//swe)

ENGINEERING AND TECHNOLOGY  -- Environmental Biotechnology -- Water Treatment (hsv//eng)

Keyword

Cell Membrane

Protein Biochemistry

Single-molecule Assays

Publication and Content Type

art (subject category)

ref (subject category)