AXOR12, a Novel Human G Protein-coupled Receptor, Activated by the Peptide KiSS-1*
- Alison I. MuirtOa,
- Larissa ChamberlaintOb,
- Nabil A. ElshourbagytOc,
- David MichalovichtOdFNe,
- Darren J. MooretOb,
- Amy CalamaritOc,
- Philip G. SzekerestOa,
- Henry M. SarautOf,
- Jon K. ChamberstOa,
- Paul MurdocktOg,
- Klaudia SteplewskitOc,
- Usman ShabontOc,
- Jane E. MillertOa,
- Susan E. MiddletontOa,
- John G. DarkertOh,
- Christopher G. C. LarminietOd,
- Shelagh WilsontOa,
- Derk J. BergsmatOc,
- Piers EmsontOb,
- Richard Faulli,
- Karen L. PhilpotttOj and
- David C. HarrisontOjFNk
- From the Departments of tOjNeurology, tOaDiscovery Biology, tOgBiotechnology and Genetics, tOhDiscovery Chemistry, and tOdBioinformatics, GlaxoSmithKline, New Frontiers Science Park, Harlow, Essex CM19 5AW, United Kingdom, the Departments of tOcBiotechnology and Genetics andtOfPulmonary Biology, GlaxoSmithKline, 709 Swedeland Road, King of Prussia, Pennsylvania, 19406, the tObNeurobiology Programme, The Babraham Institute, Babraham, Cambridge CB2 4AT, United Kingdom, and the iDepartment of Anatomy with Radiology, University of Auckland, P. O. Box 92019, Auckland, New Zealand
Abstract
A novel human G protein-coupled receptor named AXOR12, exhibiting 81% homology to the rat orphan receptor GPR54, was cloned from a human brain cDNA library. Heterologous expression of AXOR12 in mammalian cells permitted the identification of three surrogate agonist peptides, all with a common C-terminal amidated motif. High potency agonism, indicative of a cognate ligand, was evident from peptides derived from the gene KiSS-1, the expression of which prevents metastasis in melanoma cells. Quantitative reverse transcriptase-polymerase chain reaction was used to study the expression of AXOR12 and KiSS-1 in a variety of tissues. The highest levels of expression of AXOR12 mRNA were observed in brain, pituitary gland, and placenta. The highest levels ofKiSS-1 gene expression were observed in placenta and brain. A polyclonal antibody raised to the C terminus of AXOR12 was generated and used to show localization of the receptor to neurons in the cerebellum, cerebral cortex, and brainstem. The biological significance of these expression patterns and the nature of the putative cognate ligand for AXOR12 are discussed.
Footnotes
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↵* The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) .
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↵FNe Current Address: Inpharmatica Ltd., 60 Charlotte St., London W1T 2NU, United Kingdom.
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↵FNk To whom correspondence should be addressed. Tel.: 44-0-1279-622728; Fax: 44-0-1279-622371; E-mail: David_C_Harrison@gsk.com.
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Published, JBC Papers in Press, May 31, 2001, DOI 10.1074/jbc.M102743200
- Abbreviations:
- GPCR
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G protein-coupled receptor
- RT
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reverse transcriptase
- PCR
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polymerase chain reaction
- CHO
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Chinese hamster ovary
- NPFF
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neuropeptide FF
- HPLC
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high pressure liquid chromatography
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- Received March 28, 2001.
- Revision received May 23, 2001.
- The American Society for Biochemistry and Molecular Biology, Inc.
