Partial purification of the mu opioid receptor irreversibly labeled with (/sup 3/H)b-funaltrexamine

Conference · Sat Mar 01 00:00:00 EST 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5497980

Liu-Chen, L Y; Phillips, C A; Tam, S W

The mu opioid receptor in bovine striatal membranes was specifically and irreversibly labeled by incubation with 5 nM (/sup 3/H)..beta..-funaltrexamine (approx.-FNA) at 37/sup 0/C for 90 min in the presence of 100 mM NaCl. The specific and irreversible binding of (/sup 3/H)..beta..-FNA as defined by that blocked by 1 /sup +/M naloxone was about 60% of total irreversible binding. The specific irreversible binding was saturable, stereospecific, time-, temperature, and tissue-dependent. Mu opioid ligands were much more potent than delta or kappa ligands in inhibiting the specific irreversible labeling. SDS polyacrylamide gel electrophoresis of solubilized membranes in the presence of 2-mercaptoethanol yielded a major radiolabeled broad band of MW 68-97K daltons, characteristic of a glycoprotein band. This band was not observed in membranes labeled in the presence of excess unlabeled naloxone. The glycoprotein nature of the (/sup 3/H)..beta..-FNA-labeled opioid receptor was confirmed by its binding to a wheat germ agglutinin-Sepharose column and its elution with N-acetylglucosamine.

• Research Organization: E.I. Du Pont Co., Wilmington, DE
• OSTI ID: 5497980
• Report Number(s): CONF-8604222-; TRN: 86-026556
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Vol. 45:3; Conference: 70. annual meeting of the Federation of American Society for Experimental Biology, St. Louis, MO, USA, 13 Apr 1986
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
OPIUM
RECEPTORS
LABELLING
PURIFICATION
BIOCHEMICAL REACTION KINETICS
CATTLE
CELL MEMBRANES
ELECTROPHORESIS
TRACER TECHNIQUES
TRITIUM COMPOUNDS
ANALGESICS
ANIMALS
CELL CONSTITUENTS
CENTRAL NERVOUS SYSTEM DEPRESSANTS
DOMESTIC ANIMALS
DRUGS
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
NARCOTICS
ORGANIC COMPOUNDS
PROTEINS
REACTION KINETICS
RUMINANTS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques