Capturing of the free cysteine residue in the ligand-binding site by affinity labeling of the ORL1 nociceptin receptor.

@article{Matsushima2011CapturingOT,
  title={Capturing of the free cysteine residue in the ligand-binding site by affinity labeling of the ORL1 nociceptin receptor.},
  author={Ayami Matsushima and Hirokazu Nishimura and Shogo Inamine and Shiho Uemura and Yasuyuki Shimohigashi},
  journal={Bioorganic & medicinal chemistry},
  year={2011},
  volume={19 24},
  pages={
          7597-602
        }
}
All of the δ, μ, and κ opioid receptors have a free thiol group of the Cys residue in the ligand-binding site, although its functional role is not yet known. In order to examine whether or not a similar Cys is also present in the ORL1 nociceptin receptor, we attempted to identify it by affinity labeling using a specific antagonist peptide. We first treated ORL1-expressing COS-7 cell membrane preparations with the thiol-alkylation reagent N-ethylmaleimide (NEM) to perform a binding assay using… CONTINUE READING
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