Expression and purification of rat glucose transporter 1 in Pichia pastoris

Research output: Chapter in Book/Report/Conference proceedingBook chapter

Abstract

Large amounts of pure and homogenous protein are a prerequisite for several biochemical and biophysical analyses, and in particular if aiming at resolving the three-dimensional protein structure. Here we describe the production of the rat glucose transporter 1 (GLUT1), a membrane protein facilitating the transport of glucose in cells. The protein is recombinantly expressed in the yeast Pichia pastoris. It is easily maintained and large-scale protein production in shaker flasks, as commonly performed in academic research laboratories, results in relatively high yields of membrane protein. The purification protocol describes all steps needed to obtain a pure and homogenous GLUT1 protein solution, including cell growth, membrane isolation, and chromatographic purification methods.

Details

Authors
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Cell and Molecular Biology

Keywords

  • Chromatography, Glucose transporter, GLUT1, Membrane protein, Pichia pastoris, Protein expression, Protein purification
Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press
Number of pages13
Volume1713
Publication statusPublished - 2018
Publication categoryResearch
Peer-reviewedYes

Publication series

NameMethods in Molecular Biology
Volume1713
ISSN (Print)1064-3745