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

Fingerprint

Opioid Receptors
Purification
Streptavidin
Rats
Amino Acid Sequence
Brain
Amino Acids
Enkephalins
Proteins
Endorphins
Lysophosphatidylcholines
Wheat Germ Agglutinins
Deoxycholic Acid
DNA Primers
Narcotic Antagonists
Polymerase chain reaction
Molecular mass
Naloxone
Guanosine Triphosphate
GTP-Binding Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Eppler, C. M., Hulmes, J. D., Wang, J. B., Johnson, B., Corbett, M., Luthin, D. 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.

Purification and partial amino acid sequence of a μ opioid receptor from rat brain. / Eppler, C. Mark; Hulmes, Jeffrey D.; Wang, Jai Bei; Johnson, Bonita; Corbett, Marty; Luthin, David R.; Uhl, George R.; Linden, Joel.

In: Journal of Biological Chemistry, Vol. 268, No. 35, 15.12.1993, p. 26447-26451.

Research output: Contribution to journalArticle

Eppler, CM, Hulmes, JD, Wang, JB, Johnson, B, Corbett, M, Luthin, DR, Uhl, GR & Linden, J 1993, 'Purification and partial amino acid sequence of a μ opioid receptor from rat brain', Journal of Biological Chemistry, vol. 268, no. 35, pp. 26447-26451.
Eppler CM, Hulmes JD, Wang JB, Johnson B, Corbett M, Luthin DR et al. Purification and partial amino acid sequence of a μ opioid receptor from rat brain. Journal of Biological Chemistry. 1993 Dec 15;268(35):26447-26451.
Eppler, C. Mark ; Hulmes, Jeffrey D. ; Wang, Jai Bei ; Johnson, Bonita ; Corbett, Marty ; Luthin, David R. ; Uhl, George R. ; Linden, Joel. / Purification and partial amino acid sequence of a μ opioid receptor from rat brain. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 35. pp. 26447-26451.
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