Engineering a G protein-coupled receptor for structural studies: stabilization of the BLT1 receptor ground state.

ABSTRACT: Structural characterization of membrane proteins is hampered by their instability in detergent solutions. We modified here a G protein-coupled receptor, the BLT1 receptor of leukotriene B(4), to stabilize it in vitro. For this, we introduced a metal-binding site connecting the third and sixth transmembrane domains of the receptor. This modification was intended to restrain the activation-associated relative movement of these helices that results in a less stable packing in the isolated receptor. The modified receptor binds its agonist with low-affinity and can no longer trigger G protein activation, indicating that it is stabilized in its ground state conformation. Of importance, the modified BLT1 receptor displays an increased temperature-, detergent-, and time-dependent stability compare

SUBMITTER: Martin A 

PROVIDER: S-EPMC2762585 | biostudies-literature | 2009 Apr

REPOSITORIES: biostudies-literature