The Metastasis Suppressor Gene KiSS-1 Encodes Kisspeptins, the Natural Ligands of the Orphan G Protein-coupled Receptor GPR54

Masato Kotani,Michel Detheux,Ann Vandenbogaerde,David Communi,Jean-Marie Vanderwinden,Emmanuel Le Poul,Stéphane Brézillon,Richard Tyldesley,Nathalie Suarez-Huerta,Fabrice Vandeput,Cédric Blanpain,Serge N Schiffmann,Gilbert Vassart,Marc Parmentier

doi:10.1074/jbc.m104847200

Masato Kotani, Michel Detheux + Show 12 more

Open Access

https://doi.org/10.1074/jbc.m104847200

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Abstract

Natural peptides displaying agonist activity on the orphan G protein-coupled receptor GPR54 were isolated from human placenta. These 54-, 14,- and 13-amino acid peptides, with a common RF-amide C terminus, derive from the product of KiSS-1, a metastasis suppressor gene for melanoma cells, and were therefore designated kisspeptins. They bound with low nanomolar affinities to rat and human GPR54 expressed in Chinese hamster ovary K1 cells and stimulated PIP(2) hydrolysis, Ca(2+) mobilization, arachidonic acid release, ERK1/2 and p38 MAP kinase phosphorylation, and stress fiber formation but inhibited cell proliferation. Human GPR54 was highly expressed in placenta, pituitary, pancreas, and spinal cord, suggesting a role in the regulation of endocrine function. Stimulation of oxytocin secretion after kisspeptin administration to rats confirmed this hypothesis.

Highlights

G protein-coupled receptors constitute the largest family of membrane receptors [1]
These 54, 14, and 13-amino acid peptides, with a common RF-amide C terminus, derive from the product of KiSS-1, a metastasis suppressor gene for melanoma cells, and were designated kisspeptins. They bound with low nanomolar affinities to rat and human GPR54 expressed in Chinese hamster ovary K1 cells and stimulated PIP2 hydrolysis, Ca2؉ mobilization, arachidonic acid release, ERK1/2 and p38 mitogenactivated protein (MAP) kinase phosphorylation, and stress fiber formation but inhibited cell proliferation
GPR54 is an orphan G protein-coupled receptor structurally related to neuropeptide receptors and, to galanine receptors

Summary

Introduction

G protein-coupled receptors constitute the largest family of membrane receptors [1]. Over the last decade, a growing number of orphan G protein-coupled receptors have been made available by various cloning procedures such as PCR1 amplification using degenerate oligonucleotides and systematic sequencing of cDNA libraries and genomes. Natural peptides displaying agonist activity on the orphan G protein-coupled receptor GPR54 were isolated from human placenta. Purification of Natural Ligands of GPR54 —CHO-K1 cell lines expressing rat GPR54 (Fig. 2) together with G␣16 and apo-aequorin were used in a functional assay based on the luminescence of aequorin to identify natural ligands of the receptor.

Results

Conclusion