Berner, Bea, Daoutsali, Georgia, Melén, Emilia, Remper, Natália, Weszelovszká, Emma, Rothnie, Alice and Hedfalk, Kristina (2025). Successful strategies for expression and purification of ABC transporters. Biochimica et Biophysica Acta (BBA) - Biomembranes, 1867 (2),
Abstract
ATP-binding cassette (ABC) transporters are proteins responsible for active transport of various compounds, from small ions to macromolecules, across membranes. Proteins from this superfamily also pump drugs out of the cell resulting in multidrug resistance. Based on the cellular functions of ABC-transporters they are commonly associated with diseases like cancer and cystic fibrosis. To understand the molecular mechanism of this critical family of integral membrane proteins, structural characterization is a powerful tool which in turn requires successful recombinant production of stable and functional protein in good yields. In this review we have used high resolution structures of ABC transporters as a measure of successful protein production and summarized strategies for prokaryotic and eukaryotic proteins, respectively. In general, Escherichia coli is the most frequently used host for production of prokaryotic ABC transporters while human embryonic kidney 293 (HEK293) cells are the preferred host system for eukaryotic proteins. Independent of origin, at least two-steps of purification were required after solubilization in the most used detergent DDM. The purification tag was frequently cleaved off before structural characterization using cryogenic electron microscopy, or crystallization and X-ray analysis for prokaryotic proteins.
Publication DOI: | https://doi.org/10.1016/j.bbamem.2024.184401 |
---|---|
Divisions: | College of Health & Life Sciences > School of Biosciences College of Health & Life Sciences |
Funding Information: | The work presented in this study was supported by research grants from the Swedish Research Council (VR-RFI (2021-00166), Protein Production Sweden) where Kristina Hedfalk is participating with the Yeast Protein Production platform. The Aston Institute fo |
Additional Information: | Copyright © 2024 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (https://creativecommons.org/licenses/by/4.0/). |
Uncontrolled Keywords: | ABC-transporter,Recombinant production,Electron microscopy,Active transport |
Publication ISSN: | 1879-2642 |
Last Modified: | 26 Mar 2025 08:13 |
Date Deposited: | 09 Dec 2024 15:24 |
Full Text Link: | |
Related URLs: |
https://www.sci ... 005273624001329
(Publisher URL) http://www.scop ... tnerID=8YFLogxK (Scopus URL) |
PURE Output Type: | Review article |
Published Date: | 2025-02 |
Published Online Date: | 2024-11-12 |
Accepted Date: | 2024-11-08 |
Submitted Date: | 2024-07-11 |
Authors: |
Berner, Bea
Daoutsali, Georgia Melén, Emilia Remper, Natália Weszelovszká, Emma Rothnie, Alice ( ![]() Hedfalk, Kristina |