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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