Purification and partial amino acid sequence of a μ opioid receptor from rat brain

C. Mark Eppler, Jeffrey D. Hulmes, Jai Bei Wang, Bonita Johnson, Marty Corbett, David R. Luthin, George R. Uhl, Joel Linden

Research output: Contribution to journalArticle

Abstract

A rat brain opioid receptor protein was isolated by binding [∈-biotinyl-Lys32]β-endorphin to membranes, solubilizing the receptor-ligand (R·L) complex with deoxycholate-lysophosphatidylcholine and purifying on immobilized streptavidin and wheat germ agglutinin. The purified glycoprotein had a molecular mass of 60-70 kDa. Recovery of this protein was blocked by the non-selective opioid antagonist naloxone and the highly μ-selective agonist [D-Ala2,N-methyl-Phe4,Glyol5]-enkephalin but not by the highly δ-selective agonist [D-Pen2,4′-Cl-Phe4,D-Pen5]enkephalin when these compounds were added as competitors at the binding step. The 60-70-kDa receptor protein co-purified through the streptavidin column with 40-kDa protein recognized by anti-G antibodies. GTP and Na+ influenced dissociation of the solubilized R·125I-L complex and elution of the receptor and G protein from streptavidin in fashions consistent with the pharmacology of μ-opioid receptors. A 23-amino acid residue sequence from the purified receptor differs at 4 positions from a similar sequence in the murine δ-opioid receptor and is encoded within a novel rat brain cDNA isolated by polymerase chain reaction with oligonucleotide primers related to the murine δ-opioid receptor gene.

Original languageEnglish (US)
Pages (from-to)26447-26451
Number of pages5
JournalJournal of Biological Chemistry
Volume268
Issue number35
StatePublished - Dec 15 1993

ASJC Scopus subject areas

  • Biochemistry

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    Eppler, C. M., Hulmes, J. D., Wang, J. B., Johnson, B., Corbett, M., Luthin, D. R., Uhl, G. R., & Linden, J. (1993). Purification and partial amino acid sequence of a μ opioid receptor from rat brain. Journal of Biological Chemistry, 268(35), 26447-26451.