Hydrogel-assisted functional reconstitution of human P-glycoprotein (ABCB1) in giant liposomes.

ABSTRACT: This paper describes the formation of giant proteoliposomes containing P-glycoprotein (P-gp) from a solution of small proteoliposomes that had been deposited and partially dried on a film of agarose. This preparation method generated a significant fraction of giant proteoliposomes that were free of internalized vesicles, making it possible to determine the accessible liposome volume. Measuring the intensity of the fluorescent substrate rhodamine 123 (Rho123) inside and outside these giant proteoliposomes determined the concentration of transported substrates of P-gp. Fitting a kinetic model to the fluorescence data revealed the rate of passive diffusion as well as active transport by reconstituted P-gp in the membrane. This approach determined estimates for the membrane permeability coeffi

SUBMITTER: Horger KS 

PROVIDER: S-EPMC4346311 | biostudies-literature | 2015 Feb

REPOSITORIES: biostudies-literature