Novel human G-protein-coupled receptors
Section snippets
Materials and methods
Database searching. We queried the non-redundant (nr) and high-throughput genomic sequences (HTGS) databases maintained by the National Center for Biotechnology Information (NCBI) with the amino acid sequences of various GPCRs using the TBLAST algorithm [8]. Returned genomic sequences having statistically significant scores were further examined. The conceptualized protein sequences encoded by these genomic sequences were used to query the nr database to determine whether these sequences
Cloning of GPCR-encoding genes
A customized search of the HTGS database retrieved five candidate GPCR-encoding sequences clustered within a bacterial artificial chromosome (BAC) clone localized to human chromosome 11 (GenBank Accession No. AC090719). Primers were designed and used to PCR amplify the five DNA sequences. The PCR amplified products were cloned into pcDNA3 and sequenced. These clones were named GPR133, GPR134, GPR135, GPR136, and GPR137, and they encoded proteins of 350, 324, 317, 318, and 325 amino acids in
Discussion
In this report, we have described the discovery of seven novel human oGPCRs. The receptor genes are divided into two groups. GPR133, GPR134, GPR135, GPR136, and GPR137 are closely related to each other, and their presence within the same human contig suggests a tight clustering of these genes. As a group, these receptor genes most closely resembled an orphan receptor (PSGR) which shows prostate-specific localization [11], [12], [13], as well as some expression in the olfactory zone and the
Acknowledgements
This work was supported by a grant from the Canadian Institutes of Health Research.
References (32)
- et al.
Novel GPCRs and their endogenous ligands: expanding the boundaries of physiology and pharmacology
Trends Pharmacol. Sci.
(1999) - et al.
Discovery and mapping of ten novel G-protein-coupled receptor genes
Gene
(2001) - et al.
Cloning of human genes encoding novel G-protein-coupled receptors
Genomics
(1994) - et al.
Single-step method of RNA isolation by acid guanidium thiocyanate–phenol–chloroform extraction
Anal. Biochem.
(1987) - et al.
Cloning and genetic characterization of an evolutionarily conserved human olfactory receptor that is differentially expressed across species
Gene
(2001) - et al.
Isolation and characterization of a novel endogenous peptide ligand for the human APJ receptor
Biochem. Biophys. Res. Commun.
(1998) - et al.
Identification of melanin concentrating hormone (MCH) as the natural ligand for the orphan somatostatin-like receptor 1 (SLC-1)
FEBS Lett.
(1999) - et al.
Isolation and identification of melanin-concentrating hormone as the endogenous ligand of the SLC-1 receptor
Biochem. Biophys. Res. Commun.
(1999) - et al.
AXOR12, a novel human G protein-coupled receptor, activated by the peptide KiSS-1
J. Biol. Chem.
(2001) - et al.
The metastasis suppressor gene KiSS-1 encodes kisspeptins, the natural ligands of the orphan G protein-coupled receptor GPR54
J. Biol. Chem.
(2001)