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The E. coli envY gene encodes a high affinity opioid binding site
F Cabon1, J Morser, E Parmantier
1INSERM U 134, Université Pierre et Marie Curie, Hôpital de la Salpêtrière, Paris, France.
Neurochemical Research
|July 1, 1993
Summary
Researchers investigating opioid receptors inadvertently discovered that the E. coli envY gene encodes opioid binding sites. This bacterial gene, when expressed, creates sites with characteristics similar to mammalian opioid receptors.
Area of Science:
- Molecular Biology
- Neuroscience
- Genetics
Background:
- Opioid receptors are crucial for pain modulation and addiction.
- Previous research focused on mammalian systems for opioid receptor identification.
Purpose of the Study:
- To isolate a cDNA encoding an opioid receptor using a lambda ZAP cDNA library.
- To investigate the origin and function of unexpected sequences found during the isolation process.
Main Methods:
- Construction of a NG108-15 cell cDNA library in lambda ZAP vector.
- In vitro transcription-translation assay with sib selection strategy.
- DNA sequencing and homology comparison with data banks.
- Opiate binding assays on E. coli strains and transfected mammalian cells.
Main Results:
- A single phage containing a cDNA with 100% homology to E. coli envY gene was isolated.
- The envY gene was found to be responsible for opioid binding sites in E. coli.
- Expression of envY in rabbit reticulocyte lysate and mammalian cells resulted in opioid binding sites with mu and delta properties.
Conclusions:
- The E. coli envY gene encodes functional opioid binding sites.
- The envY gene represents a bacterial homolog of mammalian opioid receptors.
- This discovery opens new avenues for understanding opioid receptor evolution and function.