Fully denaturing two-dimensional electrophoresis of membrane proteins: a critical update

TL;DR

This paper reviews recent advances in two-dimensional electrophoresis techniques for membrane proteins, focusing on fully denaturing methods and comparing traditional IEF-based approaches with detergent-based zone electrophoresis.

Contribution

It provides a critical update on the performance and applications of denaturing 2-DE methods for membrane protein separation, highlighting recent progress and future prospects.

Findings

Detergent-based zone electrophoresis methods show promise for membrane protein analysis.

Traditional IEF-based 2-DE methods have limitations with hydrophobic membrane proteins.

Future developments may improve resolution and applicability of membrane protein separation techniques.

Abstract

The quality and ease of proteomics analysis depends on the performance of the analytical tools used, and thus of the performances of the protein separation tools used to deconvolute complex protein samples. Among protein samples, membrane proteins are one of the most difficult sample classes, because of their hydrophobicity and embedment in the lipid bilayers. This review deals with the recent progresses and advances made in the separation of membrane proteins by 2-DE separating only denatured proteins. Traditional 2-D methods, i.e., methods using IEF in the first dimension are compared to methods using only zone electrophoresis in both dimensions, i.e., electrophoresis in the presence of cationic or anionic detergents. The overall performances and fields of application of both types of method is critically examined, as are future prospects for this field.