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)

DOI:https://doi.org/10.1016/0024-3205(87)90492-9· OSTI ID:6501701

Simon, J; Szucs, M; Benyhe, S; Toth, G; Hepp, J; Borsodi, A; Wollemann, M; Medzihradszky, K

/sup 3/H-D-Ala/sup 2/-Leu/sup 5/-enkephalin chloromethyl ketone (/sup 3/H-DALECK) was used to label opioid receptors of frog brain membranes. The authors have previously shown that 70% of the opioid receptors are of kappa type in this preparation. The binding of /sup 3/H-DALECK was of high affinity, half maximal binding being achieved by 0.9 nM of the radioligand. The number of sites labeled was calculated to be 108 fmol/mg protein. Opioid ligands, incubated with the membranes prior to the label, inhibited /sup 3/H-DALECK binding with the following rank order: etorphine > EKC > DAGO > DALECK > DADLE. Dissociation experiments showed that 70% of the binding is irreversible. Fluorography performed after SDS-PAGE revealed specific covalent labeling of protein subunits of 90, 58 and 20 kD molecular weights. Results will be compared to those obtained in rat brain. The authors two studies demonstrate that /sup 3/H-DALECK is a useful probe for investigation the subunit structure of opioid receptors. 23 references, 5 figures, 1 table.

• OSTI ID: 6501701
• Journal Information: Life Sci.; (United States), Journal Name: Life Sci.; (United States) Vol. 41:2; ISSN LIFSA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMPHIBIANS
ANIMALS
AQUATIC ORGANISMS
AUTONOMIC NERVOUS SYSTEM AGENTS
BIOCHEMICAL REACTION KINETICS
BODY
BRAIN
CELL CONSTITUENTS
CELL MEMBRANES
CENTRAL NERVOUS SYSTEM
COMPARATIVE EVALUATIONS
COVALENCE
DATA
DRUGS
ELECTROPHORESIS
ENDORPHINS
ENKEPHALINS
EXPERIMENTAL DATA
FROGS
INFORMATION
ISOTOPE APPLICATIONS
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
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES