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Characterization of 16 human G protein-coupled receptors expressed in baculovirus-infected insect cells
- Source :
- Protein expression and purification. 44(1)
- Publication Year :
- 2005
-
Abstract
- 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.
- Subjects :
- Insect cell
biology
Gene Expression
Spodoptera
Crystallography, X-Ray
Ligands
Cell biology
Cell Line
Receptors, G-Protein-Coupled
Protein structure
Membrane protein
Cell culture
Protein A/G
biology.protein
Protein biosynthesis
Animals
Humans
Biological Assay
Receptor
Baculoviridae
hormones, hormone substitutes, and hormone antagonists
Biotechnology
G protein-coupled receptor
Protein Binding
Subjects
Details
- ISSN :
- 10465928
- Volume :
- 44
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- Protein expression and purification
- Accession number :
- edsair.doi.dedup.....c4b518afc8eb4791889d92dc23cb7a61