A novel photoaffinity ligand for the phencyclidine site of the N-methyl-D-aspartate receptor labels a Mr 120,000 polypeptide

Journal Article · Wed Apr 25 00:00:00 EDT 1990 · Journal of Biological Chemistry; (USA)

OSTI ID:7117985

Sonders, M S; Barmettler, P; Lee, J A; Kitahara, Y; Keana, J F; Weber, E ^[1]

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1. Oregon Health Sciences Univ., Portland (USA)

A radiolabeled photoaffinity ligand has been developed for the N-methyl-D-aspartate (NMDA)-preferring excitatory amino acid receptor complex. (3H)3-Azido-(5S, 10R)(+)-5-methyl-10,11-dihydro-5H- dibenzo(a,d)cyclohepten-5,10-imine (3H)3-azido-MK-801 demonstrated nearly identical affinity, density of binding sites, selectivity, pH sensitivity, and pharmacological profile in reversible binding assays with guinea pig brain homogenates to those displayed by its parent compound, MK-801. When employed in a photo-labeling protocol designed to optimize specific incorporation, (3H)3-azido-MK-801 labeled a single protein band which migrated in sodium dodecyl sulfate-polyacrylamide gels with Mr = 120,000. Incorporation of tritium into this band was completely inhibited when homogenates and (3H)3-azido-MK-801 were coincubated with 10 microM phencyclidine. These data suggest that the phencyclidine site of the NMDA receptor complex is at least in part comprised of a Mr = 120,000 polypeptide.

• OSTI ID: 7117985
• Journal Information: Journal of Biological Chemistry; (USA), Vol. 265:12; ISSN 0021-9258
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ASPARTIC ACID
RECEPTORS
PIPERIDINES
BIOCHEMICAL REACTION KINETICS
AFFINITY
BRAIN
CELL MEMBRANES
ELECTROPHORESIS
GUINEA PIGS
LIGANDS
MOLECULAR WEIGHT
POLYPEPTIDES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AMINES
AMINO ACIDS
ANIMALS
AZINES
BODY
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CENTRAL NERVOUS SYSTEM
HETEROCYCLIC COMPOUNDS
HYDROGEN COMPOUNDS
ISOTOPE APPLICATIONS
KINETICS
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
NERVOUS SYSTEM
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PEPTIDES
PROTEINS
PYRIDINES
REACTION KINETICS
RODENTS
VERTEBRATES
550201* - Biochemistry- Tracer Techniques