New publication from Poul Nissen's group - Expression strategies for structural studies of eukaryotic membrane proteins

In collaboration with Bjørn Pedersen's group, Aarhus University. Integral membrane proteins in eukaryotes are central to various cellular processes and key targets in structural biology, biotechnology and drug development. However, the number of available structures for eukaryotic membrane protein belies their physiological importance. Recently, the number of available eukaryotic membrane protein structures has been steadily increasing due to the development of novel strategies in construct design, expression and structure determination. Here, we examine the major expression systems exploited for eukaryotic membrane proteins. Additionally we strive to tabulate and describe the recent expression strategies in eukaryotic membrane protein structural biology. We find that a majority of targets have been expressed in advanced host systems and modified from their wild-type form with distinct focus on conformation and thermostabilisation. However, strategies for native protein purification should also be considered where possible, particularly in light of the recent advances in single particle cryo electron microscopy.

2016.06.29 | Karen Bech

(a) Purification strategy, (b) experimental methods used for structure determination, and (c) crystallisation methods used for eukaryotic membrane proteins over the last 2 years. Inset summarises the breakdown of usage for the polyHis, FLAG and StrepII entries in (a), respectively. Total values for each entry are indicated over the respective bar.


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