Skip to main content

Lund University Publications

LUND UNIVERSITY LIBRARIES

The rapid “teabag” method for high-end purification of membrane proteins

Hering, Jenny ; Missel, Julie Winkel ; Zhang, Liying ; Gunnarsson, Anders ; Castaldo, Marie ; Pedersen, Per Amstrup ; Ek, Margareta ; Gourdon, Pontus LU and Snijder, Harm Jan (2020) In Scientific Reports 10(1).
Abstract

Overproduction and purification of membrane proteins are generally challenging and time-consuming procedures due to low expression levels, misfolding, and low stability once extracted from the membrane. Reducing processing steps and shortening the timespan for purification represent attractive approaches to overcome some of these challenges. We have therefore compared a fast “teabag” purification method with conventional purification for five different membrane proteins (MraY, AQP10, ClC-1, PAR2 and KCC2). Notably, this new approach reduces the purification time significantly, and the quality of the purified membrane proteins is equal to or exceeds conventional methods as assessed by size exclusion chromatography, SDS-PAGE and... (More)

Overproduction and purification of membrane proteins are generally challenging and time-consuming procedures due to low expression levels, misfolding, and low stability once extracted from the membrane. Reducing processing steps and shortening the timespan for purification represent attractive approaches to overcome some of these challenges. We have therefore compared a fast “teabag” purification method with conventional purification for five different membrane proteins (MraY, AQP10, ClC-1, PAR2 and KCC2). Notably, this new approach reduces the purification time significantly, and the quality of the purified membrane proteins is equal to or exceeds conventional methods as assessed by size exclusion chromatography, SDS-PAGE and downstream applications such as ITC, crystallization and cryo-EM. Furthermore, the method is scalable, applicable to a range of affinity resins and allows for parallelization. Consequently, the technique has the potential to substantially simplify purification efforts of membrane proteins in basic and applied sciences.

(Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Scientific Reports
volume
10
issue
1
article number
16167
publisher
Nature Publishing Group
external identifiers
  • pmid:32999380
  • scopus:85091717292
ISSN
2045-2322
DOI
10.1038/s41598-020-73285-9
language
English
LU publication?
yes
id
45671857-27b0-4846-9061-d211688de0c8
date added to LUP
2020-10-22 15:11:25
date last changed
2024-09-19 08:19:29
@article{45671857-27b0-4846-9061-d211688de0c8,
  abstract     = {{<p>Overproduction and purification of membrane proteins are generally challenging and time-consuming procedures due to low expression levels, misfolding, and low stability once extracted from the membrane. Reducing processing steps and shortening the timespan for purification represent attractive approaches to overcome some of these challenges. We have therefore compared a fast “teabag” purification method with conventional purification for five different membrane proteins (MraY, AQP10, ClC-1, PAR2 and KCC2). Notably, this new approach reduces the purification time significantly, and the quality of the purified membrane proteins is equal to or exceeds conventional methods as assessed by size exclusion chromatography, SDS-PAGE and downstream applications such as ITC, crystallization and cryo-EM. Furthermore, the method is scalable, applicable to a range of affinity resins and allows for parallelization. Consequently, the technique has the potential to substantially simplify purification efforts of membrane proteins in basic and applied sciences.</p>}},
  author       = {{Hering, Jenny and Missel, Julie Winkel and Zhang, Liying and Gunnarsson, Anders and Castaldo, Marie and Pedersen, Per Amstrup and Ek, Margareta and Gourdon, Pontus and Snijder, Harm Jan}},
  issn         = {{2045-2322}},
  language     = {{eng}},
  number       = {{1}},
  publisher    = {{Nature Publishing Group}},
  series       = {{Scientific Reports}},
  title        = {{The rapid “teabag” method for high-end purification of membrane proteins}},
  url          = {{http://dx.doi.org/10.1038/s41598-020-73285-9}},
  doi          = {{10.1038/s41598-020-73285-9}},
  volume       = {{10}},
  year         = {{2020}},
}