Author:
Hering Jenny,Missel Julie Winkel,Zhang Liying,Gunnarsson Anders,Castaldo Marie,Pedersen Per Amstrup,Ek Margareta,Gourdon Pontus,Snijder Harm Jan
Abstract
AbstractOverproduction 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.
Funder
Horizon 2020 Framework Programme
China Scholarship Council
Knut och Alice Wallenbergs Stiftelse
Vetenskapsrådet
NordForsk
Lundbeckfonden
The Independent Research Fund Denmark
Publisher
Springer Science and Business Media LLC
Cited by
4 articles.
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