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Expression and characterization of short analogs of the Adenosine 2A Receptor
Author(s) -
Machlus Kellie Rae,
Kreidler Jennifer,
Robinson Clifford R.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a917-b
Subject(s) - g protein coupled receptor , folding (dsp implementation) , adenosine a2a receptor , integral membrane protein , chemistry , helix (gastropod) , transmembrane domain , receptor , transmembrane protein , membrane protein , computational biology , adenosine receptor , microbiology and biotechnology , biochemistry , membrane , biophysics , biology , agonist , ecology , snail , electrical engineering , engineering
Although much is known about the structure and corresponding function of soluble proteins, the folding mechanisms and pathways of G protein‐coupled receptors (GPCRs) are poorly understood. Our goal is to understand the specific transmembrane interactions that stabilize integral membrane proteins, and to develop a model for GPCR folding. In this investigation, the Adenosine 2A Receptor (A 2A R) was studied as a model GPCR. Like all GPCRs, A 2A R contains seven membrane‐spanning alpha‐helical domains. Recombinant DNA methods were used to create a mini‐gene of the A 2A receptor that contains trans‐membrane helices two and three, and another containing five and six. Using the pET31b(+) system, fusion constructs of these peptides have been sufficiently over‐expressed in E. coli , and purified by Ni‐Chelate chromatography. Currently, studies are underway to characterize the structure of the two‐helix fragments, and measure its interactions with other trans‐membrane helices from A 2A R. Ultimately, we are using these and similar constructs to learn about the assembly pathway and structure of A 2A R. This project was funded by HHMI and the NIH.