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 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.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Hering J,Missel JW,Zhang L,Gunnarsson A,Castaldo M,Pedersen PA,Ek M,Gourdon P,Snijder HJdoi
10.1038/s41598-020-73285-9subject
Has Abstractpub_date
2020-09-30 00:00:00pages
16167issue
1issn
2045-2322pii
10.1038/s41598-020-73285-9journal_volume
10pub_type
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