Hamster Melatonin Receptors: Cloning and Binding Characterization of MT1 and Attempt to Clone MT2
2018
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
platypusMT1, 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
hamsterMT1 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(
Phodopussungorus) expresses a non-functional MT2. We observed that unlike this
hamster, the European
hamster(Cricetus cricetus) does not have a
stop codonin 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
hamsterMT2 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.
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