Reconstitution and transport analysis of eukaryotic transporters in the post-genomic era

Hiroshi Omote, Yoshinori Moriyama

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)


Measuring transport activity through reconstituted proteoliposomes is a key technique to resolve numerous problems found in the traditional methods. The system includes overexpression, purification, and reconstitution of transporters. Mixing of purified transporter with lipid and dilution below the critical micelle concentration result in rapid generation of proteoliposomes. Incubation of proteoliposomes in the presence of a driving force initiates substrate uptake. After starting the reaction, samples are passed through a gel filtration column to separate proteoliposomes from the reaction mixture. Here, we describe step-by-step procedures for such reconstitution assays.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Number of pages10
Publication statusPublished - 2018

Publication series

NameMethods in Molecular Biology
ISSN (Print)1064-3745


  • Driving force
  • Kinetics
  • Proteoliposome
  • Purification
  • Reconstitution
  • Transporter

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics


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