Mutation W284L of the human delta opioid receptor reveals agonist specific receptor conformations for G protein activation

Yoshiaki Hosohata, Eva V. Varga, Dagmar Stropova, Xiaoping Li, Richard J. Knapp, Victor J. Hruby, Kenner C. Rice, Hiroshi Nagase, William R. Roeske, Henry I. Yamamura

Research output: Contribution to journalArticlepeer-review

17 Scopus citations


Intrinsic activities of different δ opioid agonists were determined in a [ 35 S]GTPγS binding assay using cell membranes from Chinese hamster ovary (CHO) cells stably expressing the wild type (hDOR/CHO) or W284L mutant human delta opioid receptor (W284L/CHO). Agonist binding affinities were regulated more robustly by sodium and guanine nucleotide in W284L/CHO than in hDOR/CHO cell membranes. The W284L mutation selectively reduced the affinity of SNC 80 while having moderate effect ((-) TAN 67) or no effect (DPDPE) on the affinities of other δ selective agonists. The mutation had opposite effects on the intrinsic activities of agonists belonging to different chemical classes. The effects of the mutation on agonist affinities and potencies were independent from its effects on the intrinsic activities of the agonists. Maximal stimulation of [ 35 S]GTPγS binding by SNC 80 was 2-fold higher in W284L mutant cell membranes than in wild type hDOR/CHO cell membranes, despite lower receptor expression levels in the W284L/CHO cells. The binding affinity of SNC 80 however, was significantly reduced (15-fold and 30-fold in the absence or presence of sodium+GDP respectively) in W284L/CHO cell membranes relative to wild type hDOR/CHO membranes. Conversely, the E max of (-)TAN 67 in the [ 35 S]GTPγS binding assay was markedly reduced (0.6-fold of that of the wild type) with only a slight (6-fold) reduction in its binding affinity. The affinity and intrinsic activity of DPDPE on the other hand remained unchanged at the W284L mutant hDOR. The mutation had similar effects on the affinities potencies and intrinsic activities of (-)TAN 67 and SB 219825. The results indicate that δ opioid agonists of different chemical classes use specific conformations for G protein activation.

Original languageEnglish (US)
Pages (from-to)2233-2242
Number of pages10
JournalLife Sciences
Issue number19-20
StatePublished - Apr 6 2001


  • (-)TAN 67
  • Human delta opioid receptor
  • Intrinsic activity
  • SB 219825
  • SNC 80
  • W284L point mutation
  • [ S]GTPγS binding

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)


Dive into the research topics of 'Mutation W284L of the human delta opioid receptor reveals agonist specific receptor conformations for G protein activation'. Together they form a unique fingerprint.

Cite this