Liposome reconstitution and transport assay for recombinant transporters.

Zachary Lee Johnson, Seok-Yong Lee

Methods in enzymology 2015

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Secondary active transporters are responsible for the cellular uptake of many biologically important molecules, including neurotransmitters, nutrients, and drugs. Because of their physiological and clinical importance, a method for assessing their transport activity in vitro is necessary to gain a better understanding of how these transporters function at the molecular level. In this chapter, we describe a protocol for reconstituting the concentrative nucleoside transporter from Vibrio cholerae into proteoliposomes. We then describe a radiolabeled substrate uptake assay that can be used to functionally characterize the transporter. These methods are relatively common and can be applied to other secondary active transporters, with or without some modification. © 2015 Elsevier Inc. All rights reserved.

Citation

Zachary Lee Johnson, Seok-Yong Lee. Liposome reconstitution and transport assay for recombinant transporters. Methods in enzymology. 2015;556:373-83