title: Engineering a soluble parathyroid hormone GPCR mimetic.
abstract: We designed and characterized a soluble mimic of the parathyroid hormone (PTH) receptor (PTH1R) that incorporates the N-terminus and third extracellular loop of PTH1R, important for ligand binding. The engineered receptor (PTH1R-NE3) was conceived to enable easy production and the use of standard biochemical and biophysical assays for the screening of competitive antagonists of PTH. We show that PTH1R-NE3 is folded, thermodynamically stable and selectively binds PTH. We also demonstrate the utility of our mimic by identifying a small molecule that competes with PTH in our PTH1R-NE3-based fluorescence polarization assay. Antagonists to PTH1R, a transmembrane protein belonging to the class B G-protein coupled receptor family, may provide new therapeutic options for calcium metabolism diseases like humoral hypercalcemia of malignancy.
authors.affiliation: Department of Chemistry, Dartmouth College, Hanover, New Hampshire, 03755.
authors.forename: Christopher O
authors.initials: CO
authors.lastname: Audu
authors.forename: Jessica J
authors.initials: JJ
authors.lastname: Plati
authors.forename: Maria
authors.initials: M
authors.lastname: Pellegrini
authors.forename: Dale F
authors.initials: DF
authors.lastname: Mierke
mesh_terms: D006801:Humans; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D015202:Protein Engineering / Q000379:methods*; D044168:Receptor, Parathyroid Hormone, Type 1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012995:Solubility
chemical_list: D044168:Receptor, Parathyroid Hormone, Type 1; D011993:Recombinant Fusion Proteins
keywords: GPCR; NMR; drug development; drug screening; fluorescence polarization; photoaffinity crosslinking; receptor mimetic