Molecular characterization of cardiac. beta. -adrenergic receptors

Thesis/Dissertation · Thu Dec 31 23:00:00 EST 1987

OSTI ID:7147208

Robinson, D A

A rat cardiac cDNA clone and a rat genomic clone encoding the {beta}{sub 2}-adrenergic receptor were isolated and characterized by restriction mapping and automated DNA sequence analysis. The rat cardiac {beta}{sub 2}-receptor consists of 418 amino acids with a calculated molecular weight of 46,890 Da. In order to characterize the rat cardiac {beta}{sub 2}-receptor in more detail, B-82 cells, a murine fibroblast cell line lacking intrinsic {beta}-adrenergic receptors, were transfected with the gene encoding the rat {beta}{sub 2}-receptor, using a vector that allows stable, steroid-inducible expression of mammalian genes. Membranes prepared from transfected cells exhibited specific binding of the {beta}-receptor antagonist ({sup 125}I)iodocyanopindolol (ICYP) that was saturable and of high affinity. Displacement of ICYP binding to rat cardiac receptors by {beta}-adrenergic ligands was stereoselective and exhibited a rank order of potency for agonists expected for a {beta}{sub 2}-adrenergic receptor subtype. Expressed {beta}{sub 2}-receptors photoaffinity labeled with ({sup 125}I)iodocyanopindolol diazirine (ICYP-D) displayed as SDS-PAGE apparent molecular mass of 68,000 suggesting that the receptor is glycosylated 30% by weight. B-82 cells transfected with the rat {beta}{sub 2}-receptor gene exhibited a dose-dependent increase in intracellular levels of cyclic AMP in response to stimulation by isoproterenol.

• Research Organization: State Univ. of New York, Stony Brook, NY (USA). Coll. of Engineering
• OSTI ID: 7147208
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
AMP
ANIMAL CELLS
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CARDIOVASCULAR SYSTEM
CELL CONSTITUENTS
CELL MEMBRANES
CLONING
CONNECTIVE TISSUE CELLS
DAYS LIVING RADIOISOTOPES
DNA HYBRIDIZATION
DNA SEQUENCING
DNA-CLONING
DOSE-RESPONSE RELATIONSHIPS
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
FIBROBLASTS
GENE REGULATION
GENES
HEART
HYBRIDIZATION
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LIGANDS
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
MICE
MOLECULAR BIOLOGY
MOLECULAR STRUCTURE
MOLECULAR WEIGHT
NUCLEI
NUCLEOTIDES
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RADIOISOTOPES
RATS
REACTION KINETICS
RECEPTORS
RODENTS
SOMATIC CELLS
STRUCTURAL CHEMICAL ANALYSIS
SYMPATHOLYTICS
SYMPATHOMIMETICS
TRACER TECHNIQUES
VERTEBRATES