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

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.

journal_name

Int J Mol Sci

journal_title

International journal of molecular sciences

authors

Gautier C,Dufour E,Dupré C,Lizzo G,Caignard S,Riest-Fery I,Brasseur C,Legros C,Delagrange P,Nosjean O,Simonneaux V,Boutin JA,Guenin SP

doi

10.3390/ijms19071957

subject

Has Abstract

pub_date

2018-07-04 00:00:00

issue

issn

1422-0067

pii

ijms19071957

journal_volume

pub_type

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