Reconstitutively active G protein-coupled receptors purified from baculovirus-infected insect cells

摘要: Abstract The turkey beta-adrenergic receptor (beta-AR), the m1 and m2 forms of human muscarinic cholingeric (MAChR) several other mutant wild-type G protein-coupled receptors were produced in insect Sf9 cells by infection with recombinant baculoviruses. Maximal expression for most was 5-30 pmol receptor/mg protein (2-15 nmol/liter culture). displayed typical ligand binding characteristics. beta-AR glycosylated; electrophoretic behavior two MAChRs also suggested glycosylation. stimulated endogenous adenylyl cyclase response to agonists. both purified coreconstituted various proteins phospholipid vesicles. catalyzed agonist-dependent activation Gs guanosine 5'-O-(thiotriphosphate) (GTP gamma S) same efficiency as did natural beta-AR. MAChR efficiently GTP S Go recently identified Gz (Gx). Gj,1 Gj,3 weakly. Activation these ml much less efficient, consistent its known selectivity pertussis toxin-insensitive ("Gp") that have not yet been isolated. characteristically reduction disulfides. These results demonstrate general utility baculovirus system production large quantities native receptors.