title: Genetic Variations in the Human G Protein-coupled Receptor Class C, Group 6, Member A (GPRC6A) Control Cell Surface Expression and Function.
abstract: GPRC6A is a G protein-coupled receptor activated by l-amino acids, which, based on analyses of knock-out mice, has been suggested to have physiological functions in metabolism and testicular function. The human ortholog is, however, mostly retained intracellularly in contrast to the cell surface-expressed murine and goldfish orthologs. The latter orthologs are Gq-coupled and lead to intracellular accumulation of inositol phosphates and calcium release. In the present study we cloned the bonobo chimpanzee GPRC6A receptor, which is 99% identical to the human receptor, and show that it is cell surface-expressed and functional. By analyses of chimeric human/mouse and human/bonobo receptors, bonobo receptor mutants, and the single nucleotide polymorphism database at NCBI, we identify an insertion/deletion variation in the third intracellular loop responsible for the intracellular retention and lack of function of the human ortholog. Genetic analyses of the 1000 genome database and the Inter99 cohort of 6,000 Danes establish the distribution of genotypes among ethnic groups, showing that the cell surface-expressed and functional variant is much more prevalent in the African population than in European and Asian populations and that this variant is partly linked with a stop codon early in the receptor sequence (rs6907580, amino acid position 57). In conclusion, our data solve a more than decade-old question of why the cloned human GPRC6A receptor is not cell surface-expressed and functional and provide a genetic framework to study human phenotypic traits in large genome sequencing projects linked with physiological measurement and biomarkers.
authors.affiliation: From the Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Stine
authors.initials: S
authors.lastname: Jørgensen
authors.affiliation: the Novo Nordisk Foundation Center for Basic Metabolic Research, Section of Metabolic Genetics, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200 Copenhagen, Denmark.
authors.forename: Christian Theil
authors.identifier: 0000-0002-8634-4781
authors.initials: CT
authors.lastname: Have
authors.affiliation: From the Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Christina Rye
authors.initials: CR
authors.lastname: Underwood
authors.affiliation: From the Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Lars Dan
authors.initials: LD
authors.lastname: Johansen
authors.affiliation: From the Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Petrine
authors.identifier: 0000-0002-5455-8013
authors.initials: P
authors.lastname: Wellendorph
authors.affiliation: the Novo Nordisk Foundation Center for Basic Metabolic Research, Section of Metabolic Genetics, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200 Copenhagen, Denmark.
authors.forename: Anette Prior
authors.identifier: 0000-0003-4580-1576
authors.initials: AP
authors.lastname: Gjesing
authors.affiliation: From the Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark.
authors.forename: Christinna V
authors.initials: CV
authors.lastname: Jørgensen
authors.affiliation: BGI-Shenzhen, Shenzhen 518083, China.
authors.forename: Shi
authors.initials: S
authors.lastname: Quan
authors.affiliation: BGI-Shenzhen, Shenzhen 518083, China.
authors.forename: Gao
authors.initials: G
authors.lastname: Rui
authors.affiliation: the Laboratory of Molecular and Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980-8578, Japan.|the Japan Science and Technology Agency, Precursory Research for Embryonic Science and Technology, Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Asuka
authors.initials: A
authors.lastname: Inoue
authors.affiliation: the Research Centre for Prevention and Health, Capital Region of Denmark, DK-2600 Glostrup, Denmark.|the Department of Clinical Experimental Research, Rigshospitalet, DK-2100 Copenhagen, Denmark, and.|the Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200 Copenhagen, Denmark.
authors.forename: Allan
authors.identifier: 0000-0002-0994-0184
authors.initials: A
authors.lastname: Linneberg
authors.affiliation: the Novo Nordisk Foundation Center for Basic Metabolic Research, Section of Metabolic Genetics, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200 Copenhagen, Denmark.
authors.forename: Niels
authors.identifier: 0000-0001-5526-1070
authors.initials: N
authors.lastname: Grarup
authors.affiliation: BGI-Shenzhen, Shenzhen 518083, China.
authors.forename: Wang
authors.initials: W
authors.lastname: Jun
authors.affiliation: the Novo Nordisk Foundation Center for Basic Metabolic Research, Section of Metabolic Genetics, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200 Copenhagen, Denmark.
authors.forename: Oluf
authors.initials: O
authors.lastname: Pedersen
authors.affiliation: the Novo Nordisk Foundation Center for Basic Metabolic Research, Section of Metabolic Genetics, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2200 Copenhagen, Denmark.
authors.forename: Torben
authors.initials: T
authors.lastname: Hansen
authors.affiliation: From the Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, DK-2100 Copenhagen, Denmark, hbo@sund.ku.dk.
authors.forename: Hans
authors.identifier: 0000-0001-9495-7388
authors.initials: H
authors.lastname: Bräuner-Osborne
mesh_terms: D000818:Animals; D002460:Cell Line; D005786:Gene Expression Regulation; D006801:Humans; D054643:INDEL Mutation; D051379:Mice; D017433:Protein Structure, Secondary; D043562:Receptors, G-Protein-Coupled / Q000096:biosynthesis / Q000235:genetics
chemical_list: C488584:GPRC6A protein, human; C515750:GPRC6A protein, mouse; D043562:Receptors, G-Protein-Coupled
keywords: 7-helix receptor; G protein-coupled receptor (GPCR); GPRC6A receptor; cell surface receptor; human genetics; molecular pharmacology; third intracellular loop