Partial purification of the mu opioid receptor irreversibly labeled with (/sup 3/H)b-funaltrexamine
Abstract
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.
- Authors:
- Publication Date:
- Research Org.:
- E.I. Du Pont Co., Wilmington, DE
- OSTI Identifier:
- 5497980
- Report Number(s):
- CONF-8604222-
Journal ID: CODEN: FEPRA; TRN: 86-026556
- Resource Type:
- Conference
- Journal Name:
- Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
- Additional Journal Information:
- Journal Volume: 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
- Subject:
- 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
Citation Formats
Liu-Chen, L Y, Phillips, C A, and Tam, S W. Partial purification of the mu opioid receptor irreversibly labeled with (/sup 3/H)b-funaltrexamine. United States: N. p., 1986.
Web.
Liu-Chen, L Y, Phillips, C A, & Tam, S W. Partial purification of the mu opioid receptor irreversibly labeled with (/sup 3/H)b-funaltrexamine. United States.
Liu-Chen, L Y, Phillips, C A, and Tam, S W. 1986.
"Partial purification of the mu opioid receptor irreversibly labeled with (/sup 3/H)b-funaltrexamine". United States.
@article{osti_5497980,
title = {Partial purification of the mu opioid receptor irreversibly labeled with (/sup 3/H)b-funaltrexamine},
author = {Liu-Chen, L Y and Phillips, C A and Tam, S W},
abstractNote = {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.},
doi = {},
url = {https://www.osti.gov/biblio/5497980},
journal = {Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)},
number = ,
volume = 45:3,
place = {United States},
year = {Sat Mar 01 00:00:00 EST 1986},
month = {Sat Mar 01 00:00:00 EST 1986}
}