Characterization of 16 human G protein-coupled receptors expressed in baculovirus-infected insect cells

Understanding the three-dimensional structure of G protein-coupled receptors (GPCRs) has been limited by the technical challenges associated with expression, purification, and crystallization of membrane proteins, and their low abundance in native tissue. In the first large-scale comparative study o...

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Bibliographic Details
Published in:Protein expression and purification 2005-11, Vol.44 (1), p.65-74
Main Authors: Akermoun, Malika, Koglin, Markus, Zvalova-Iooss, Darina, Folschweiller, Nicolas, Dowell, Simon J., Gearing, Katy L.
Format: Article
Language:English
Subjects:
Animals
Baculoviridae
Baculovirus
Biological Assay - methods
Bmax
Cell Line
Crystallography, X-Ray - methods
Expression
G protein-coupled receptor
Gene Expression - genetics
Humans
Ligands
Protein Binding
Protein structure
Receptors, G-Protein-Coupled - chemistry
Receptors, G-Protein-Coupled - genetics
Receptors, G-Protein-Coupled - metabolism
Spodoptera
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Summary:Understanding the three-dimensional structure of G protein-coupled receptors (GPCRs) has been limited by the technical challenges associated with expression, purification, and crystallization of membrane proteins, and their low abundance in native tissue. In the first large-scale comparative study of GPCR protein production using recombinant baculovirus, we report the characterization of 16 human receptors. The GPCRs were produced in three insect cell lines and functional protein levels monitored over 72 h using radioligand binding assays. Different GPCRs exhibited widely different expression levels, ranging from less than 1 pmol receptor/mg protein to more than 250 pmol/mg. No single set of conditions was suitable for all GPCRs, and large differences were seen for the expression of individual GPCRs in different cell lines. Closely related GPCRs did not share similar expression profiles; however, high expression (greater than 20 pmol/mg) was achieved for over half the GPCRs in our study. Overall, the levels of protein production compared favourably to other published systems.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2005.04.016