Structure of signaling-competent neurotensin receptor 1 obtained by directed evolution in Escherichia coli.

ABSTRACT: Crystallography has advanced our understanding of G protein-coupled receptors, but low expression levels and instability in solution have limited structural insights to very few selected members of this large protein family. Using neurotensin receptor 1 (NTR1) as a proof of principle, we show that two directed evolution technologies that we recently developed have the potential to overcome these problems. We purified three neurotensin-bound NTR1 variants from Escherichia coli and determined their X-ray structures at up to 2.75 Å resolution using vapor diffusion crystallization experiments. A crystallized construct was pharmacologically characterized and exhibited ligand-dependent signaling, internalization, and wild-type-like agonist and antagonist affinities. Our structures are fully cons

SUBMITTER: Egloff P 

PROVIDER: S-EPMC3926081 | biostudies-literature | 2014 Feb

REPOSITORIES: biostudies-literature