Back to Search Start Over

Hamster Melatonin Receptors: Cloning and Binding Characterization of MT1 and Attempt to Clone MT2

Authors :
Célia Gautier
Emilie Dufour
Clémence Dupré
Giulia Lizzo
Sarah Caignard
Isabelle Riest-Fery
Chantal Brasseur
Céline Legros
Philippe Delagrange
Olivier Nosjean
Valérie Simonneaux
Jean A. Boutin
Sophie-Pénélope Guenin
Source :
International Journal of Molecular Sciences, Vol 19, Iss 7, p 1957 (2018)
Publication Year :
2018
Publisher :
MDPI AG, 2018.

Abstract

For many years, it was of interest to identify the sequences encoding the two melatonin receptors (MT1 and MT2) from various species. After publishing the basic molecular characterization of the human, rat, mouse, sheep, and platypus MT1, MT2, or Mel1c receptors, we began cloning the genes from other animals, such as birds, bats, and vipers. The goal was to advance the receptor crystallization, which could greatly contribute the understanding of the sequence/stability relationship. European hamster MT1 receptor was cloned for the first time from this gender, was expressed in stable form in cells, and its binding characterized with a sample of 19 melatonin ligands. Siberian hamster (Phodopus sungorus) expresses a non-functional MT2. We observed that unlike this hamster, the European hamster (Cricetus cricetus) does not have a stop codon in the MT2 sequence. Thus, we undertook the tedious task of cloning the MT2 receptor. We partially succeeded, sequencing the complete exon 2 and a fragment of exon 1 (from putative amino acids 12 to 38 and 77 to 323), after several years of efforts. In order to show that the protein parts we cloned were capable to sustain some binding capacities, we designed a chimeric MT2 receptor using a consensus sequence to replace the unknown amino acids, based on other small rodent MT2 sequences. This chimeric construct could bind melatonin in the nanomolar range. This work is meant to be the basis for attempts from other laboratories of the community to determine the complete natural sequence of the European hamster MT2 receptor. The present work is the first to show that, among the hamsters, if the Siberian is a natural knockout for MT2, the European one is not.

Details

Language :
English
ISSN :
14220067 and 19071957
Volume :
19
Issue :
7
Database :
Directory of Open Access Journals
Journal :
International Journal of Molecular Sciences
Publication Type :
Academic Journal
Accession number :
edsdoj.3a1d8bb090f143ef935822001e658ada
Document Type :
article
Full Text :
https://doi.org/10.3390/ijms19071957