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 compared with the wild-type receptor. These data indicate that stabilizing the ground state of this GPCR by limiting the activation-associated movements of the transmembrane helices is a way to increase its stability in detergent solutions; this could represent a forward step on the way of its crystallization.
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Protein Science, Wiley, 2009, 18 (4), pp.727-34. 〈10.1002/pro.55〉
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Aimée Martin, Marjorie Damian, Michel Laguerre, Joseph Parello, Bernard Pucci, et al.. Engineering a G protein-coupled receptor for structural studies: stabilization of the BLT1 receptor ground state.. Protein Science, Wiley, 2009, 18 (4), pp.727-34. 〈10.1002/pro.55〉. 〈inserm-00868966〉

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