Expression Systems for GPCRs
Vector
Advantages
Cell-Free Systems
E. coli; wheat germ Rapid, PCR applicable
Prokaryotes — Bacteria
E. coli MB Rapid, inexpensive, scaleable
E. coli IB
Halobacterium salinarium
Lactococcus lactis
Eukaryotes — Yeasts
Saccharomyces cerevisiae
Schizosaccharomyces pombe Hansenula polymorpha Pichia pastoris
Rapid, inexpensive, high yields, easy scale-up
Rapid, scaleable, colorimetric expression
Rapid, scaleable, inexpensive
Relatively easy to use, high biomass, scaleable Relatively inexpensive Inducible
High yields
Eukaryotes — Insect Cells
Baculovirus Mammalian-like glycosylation
Drosophila S2
GFP fusion
Eukaryotes — Mammalian
Transient Native processing
Stable
Inducible
Disadvantages
Expensive, for soluble proteins
Toxic to cells, no post-translational modifications Low yields Requires refolding
Cloning complicated, no post-translational modifications, fusion proteins No post-translation modifications
Hyperglycosylation, clone selection, thick cell wall Thick cell wall
Low yields
Thick cell wall
More expensive, virus stock prep slow Time-consuming
Expensive
Time-consuming, low yields
Target GPCR Ref.
Human A2aR Rat NTR Human 5HT1AR
Human LPTHR1 Human LGLP-1 Human leukotriene BLT1 Human muscarinic M1R Human 5HT2CR Yeast Ste2R
No GPCRs ABC transporters
Yeast Ste2R Dopamine D1AR
Dopamine D2R
Chemokine receptors Human ß2 AR 5HT5A receptor
Human NKIR Human ß2 AR
Human opioid receptor
Muscarinic M1, M4R Human ß2 AR
15 5
26 26
34 34
40 40
49 52
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