Fusion partner toolchest for the stabilization and crystallization of G protein-coupled receptors. Structure 2012 Jun 06;20(6):967-76
Date
06/12/2012Pubmed ID
22681902Pubmed Central ID
PMC3375611DOI
10.1016/j.str.2012.04.010Scopus ID
2-s2.0-84861984672 (requires institutional sign-in at Scopus site) 321 CitationsAbstract
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.
Author List
Chun E, Thompson AA, Liu W, Roth CB, Griffith MT, Katritch V, Kunken J, Xu F, Cherezov V, Hanson MA, Stevens RCAuthor
Wei Liu PhD Associate Professor in the Pharmacology and Toxicology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Amino Acid SequenceAnimals
Cell Line
Chromatography, Gel
Cloning, Molecular
Crystallization
Crystallography, X-Ray
Cytochromes b
Humans
Molecular Sequence Data
Muramidase
Protein Stability
Receptor, Adenosine A2A
Receptors, G-Protein-Coupled
Recombinant Fusion Proteins