Use of anionic denaturing detergents to purify insoluble proteins after overexpression.

ABSTRACT:

Background

Many proteins form insoluble protein aggregates, called "inclusion bodies", when overexpressed in E. coli. This is the biggest obstacle in biotechnology. Ever since the reversible denaturation of proteins by chaotropic agents such as urea or guanidinium hydrochloride had been shown, these compounds were predominantly used to dissolve inclusion bodies. Other denaturants exist but have received much less attention in protein purification. While the anionic, denaturing detergent sodiumdodecylsulphate (SDS) is used extensively in analytical SDS-PAGE, it has rarely been used in preparative purification.

Results

Here we present a simple and versatile method to purify insoluble, hexahistidine-tagged proteins under denaturing conditions. It is based on dissolution of over

SUBMITTER: Schlager B 

PROVIDER: S-EPMC3536628 | biostudies-literature | 2012 Dec

REPOSITORIES: biostudies-literature