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Fusion partner toolchest for the stabilization and crystallization of G protein-coupled receptors
Authors:Chun Eugene  Thompson Aaron A  Liu Wei  Roth Christopher B  Griffith Mark T  Katritch Vsevolod  Kunken Joshua  Xu Fei  Cherezov Vadim  Hanson Michael A  Stevens Raymond C
Affiliation:Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
Abstract:Structural studies of human G protein-coupled receptors (GPCRs) have recently been accelerated through the use of a fusion partner that was inserted into the third intracellular loop. Using chimeras of the human β(2)-adrenergic and human A(2A) adenosine receptors, we present the methodology and data for the initial selection of an expanded set of fusion partners for crystallizing GPCRs. In particular, use of the thermostabilized apocytochrome b(562)RIL as a fusion partner displays certain advantages over previously utilized fusion proteins, resulting in?a significant improvement in stability and structure of GPCR-fusion constructs.
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