title: Expression of CXCR4, a G-protein-coupled receptor for CXCL12 in yeast identification of new-generation inverse agonists.
abstract: G-protein-coupled receptors (GPCR) are prime targets for therapies with small molecule-antagonists. Since yeast have GPCR triggered signaling pathways analogous to those present in mammalian cells, it is possible to express human receptors in yeast coupled to the pheromone responsive signaling cascade in variants that contain mammalian-yeast Galpha subunit chimeras. CXCR4 and CXCR4(N119S), a constitutively active mutant were expressed in yeast coupled to pheromone responsive reporter genes, HIS3, lacZ, or FUI, and tested for signaling activity. Compounds derived from T140, an inverse agonist for CXCR4, were screened for activity using yeast cells expressing CXCR4(N119S) and containing a FUS1-lacZ reporter gene. Levels of inhibition of beta-galactosidase activities triggered by constitutive activation of the pheromone response pathway that were obtained in the presence of the T140 derived compounds correlated with affinities measured in radioligand binding inhibition experiments. The yeast signaling system may provide an effective approach for screening chemokine receptor antagonists.
authors.affiliation: Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylvania, USA.
authors.forename: Barry J
authors.initials: BJ
authors.lastname: Evans
authors.forename: Zixuan
authors.initials: Z
authors.lastname: Wang
authors.forename: James R
authors.initials: JR
authors.lastname: Broach
authors.forename: Shinya
authors.initials: S
authors.lastname: Oishi
authors.forename: Nobutaka
authors.initials: N
authors.lastname: Fujii
authors.forename: Stephen C
authors.initials: SC
authors.lastname: Peiper
mesh_terms: D054377:Chemokine CXCL12 / Q000378:metabolism*; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D006801:Humans; D009842:Oligopeptides / Q000737:chemistry / Q000494:pharmacology; D010456:Peptides, Cyclic / Q000494:pharmacology; D019718:Receptors, CXCR4 / Q000819:agonists* / Q000235:genetics / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000819:agonists / Q000378:metabolism*
chemical_list: D054377:Chemokine CXCL12; D009842:Oligopeptides; D010456:Peptides, Cyclic; D019718:Receptors, CXCR4; D029701:Saccharomyces cerevisiae Proteins; C496951:cyclo(tyrosyl-arginyl-arginyl-3-(2-naphthyl)alanyl-glycyl); C116977:T140 peptide