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 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.
Akermoun
Malika
M
Gene Expression and Protein Biochemistry, GlaxoSmithKline Research and Development, Medicines Research Centre, Gunnels Wood Road, Stevenage, Hertfordshire SG1 2NY, UK.
Koglin
Markus
M
Zvalova-Iooss
Darina
D
Folschweiller
Nicolas
N
Dowell
Simon J
SJ
Gearing
Katy L
KL
D000818:Animals; D016367:Baculoviridae; D001681:Biological Assay / Q000379:methods; D002460:Cell Line; D018360:Crystallography, X-Ray / Q000379:methods; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D008024:Ligands; D011485:Protein Binding; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018411:Spodoptera
D008024:Ligands; D043562:Receptors, G-Protein-Coupled