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), Vol. 41:2
• Country of Publication: United States
• Language: English

Related Subjects

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