Expression strategies for structural studies of eukaryotic membrane proteins

Joseph A Lyons; Azadeh Shahsavar; Peter Aasted Paulsen; Bjørn Panyella Pedersen; Poul Nissen

doi:10.1016/j.sbi.2016.06.011

Expression strategies for structural studies of eukaryotic membrane proteins

Joseph A Lyons, Azadeh Shahsavar, Peter Aasted Paulsen, Bjørn Panyella Pedersen, Poul Nissen

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Abstract

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.

Original language	English
Journal	Current Opinion in Structural Biology
Volume	38
Pages (from-to)	137-144
Number of pages	8
ISSN	0959-440X
DOIs	https://doi.org/10.1016/j.sbi.2016.06.011
Publication status	Published - 1 Jun 2016

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title = "Expression strategies for structural studies of eukaryotic membrane proteins",

abstract = "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.",

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T1 - Expression strategies for structural studies of eukaryotic membrane proteins

AU - Lyons, Joseph A

AU - Shahsavar, Azadeh

AU - Paulsen, Peter Aasted

AU - Pedersen, Bjørn Panyella

AU - Nissen, Poul

PY - 2016/6/1

Y1 - 2016/6/1

N2 - 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.

AB - 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.

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Expression strategies for structural studies of eukaryotic membrane proteins

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