Skip to main content
OSTI.GOVU.S. Department of Energy Office of Scientific and Technical Information
U.S. Department of Energy
Office of Scientific and Technical Information
 

Covalent labeling of opioid receptors with /sup 3/H-D-Ala/sup 2/-Leu/sup 5/-enkephalin chloromethyl ketone I. Binding characteristics in rate brain membranes

Journal Article · · Life Sci.; (United States)
; ; ; ; ; ; ;

The chloromethyl ketone derivative of D-Ala/sup 2/-Leu/sup 5/-enkephalin was synthesized in a radioactive form, and the resulting compound (/sup 3/H-DALECK) was used to label opioid receptors. /sup 3/H-DALECK binds with high affinity, specificity and saturability to rat brain membranes. The number of sites labeled is 130 fmoles/mg protein. Unlabeled opioids inhibited the binding of /sup 3/H-DALECK; etorphine and DAGO being most potent. A 10-fold preference for mu sites over delta was seen in site-specific competition experiments; while DALECK displayed low affinity for kappa sites of rat brain DALECK irreversibly blocked a certain population of sites. Approximately 40% of /sup 3/H-DALECK binding at 15 min, and 60% at 60 min association time did not dissociate in the presence of a large excess of unlabeled DALECK and was resistant to washing. Autoradiography performed after SDS-PAGE revealed specific alkylation of proteins with molecular weights of 74, 65, 56, 43 and 34 kD. These results demonstrate the applicability of using /sup 3/H-DALECK to covalently label opioid receptors. 21 references, 4 figures, 1 table.

OSTI ID:
6501714
Journal Information:
Life Sci.; (United States), Journal Name: Life Sci.; (United States) Vol. 41:2; ISSN LIFSA
Country of Publication:
United States
Language:
English

Similar Records

Covalent labeling of opioid receptors with /sup 3/H-D-Ala/sup 2/-Leu/sup 5/-enkephalin chloromethyl ketone. II. Binding characteristics in frog brain membranes
Journal Article · Mon Jul 13 00:00:00 EDT 1987 · Life Sci.; (United States) · OSTI ID:6501701
The opioid receptors of the rat periaqueductal gray
Thesis/Dissertation · Sat Dec 31 23:00:00 EST 1988 · OSTI ID:6940386
Potent cyclic enkephalin analogues for delta opioid receptors in the rat brain
Conference · Fri Feb 28 23:00:00 EST 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) · OSTI ID:5278567

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
AUTORADIOGRAPHY
BIOCHEMICAL REACTION KINETICS
BODY
BRAIN
CELL CONSTITUENTS
CELL MEMBRANES
CENTRAL NERVOUS SYSTEM
COVALENCE
DATA
DRUGS
ELECTROPHORESIS
ENDORPHINS
ENKEPHALINS
EXPERIMENTAL DATA
INFORMATION
KETONES
KINETICS
LABELLED COMPOUNDS
LABELLING
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
NERVOUS SYSTEM
NEUROREGULATORS
NUMERICAL DATA
ORGANIC COMPOUNDS
ORGANS
PEPTIDES
POLYPEPTIDES
PROTEINS
RATS
REACTION KINETICS
RECEPTORS
RODENTS
TRITIUM COMPOUNDS
VERTEBRATES