A generic selection system for improved expression and thermostability of G protein-coupled receptors by directed evolution.

ABSTRACT: Structural and biophysical studies as well as drug screening approaches on G protein-coupled receptors (GPCRs) have been largely hampered by the poor biophysical properties and low expression yields of this largest class of integral membrane proteins. Thermostabilisation of GPCRs by introduction of stabilising mutations has been a key factor to overcome these limitations. However, labelled ligands with sufficient affinity, which are required for selective binding to the correctly folded receptor, are often not available. Here we describe a novel procedure to improve receptor expression and stability in a generic way, independent of specific ligands, by means of directed evolution in E. coli. We have engineered a homogenous fluorescent reporter assay that only detects receptors which are co

SUBMITTER: Klenk C 

PROVIDER: S-EPMC4758055 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature