Purification and molecular characterization of the cardiac beta-adrenergic receptor

Conference · Thu May 01 00:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5247190

Robinson, D A; Venter, J C; Fraser, C M

The porcine ventricle cardiac beta-adrenergic receptor (..beta..AR) is predominantly of the ..beta../sub 2/ subtype. Sucrose density gradient purified porcine ventricle membranes exhibit a high ..beta..AR density of 1 pmol per mg and a K/sub d/ for /sup 125/I-iodocyanopindolol of 180 pM. Digitonin solubilized receptor exhibits ligand binding characteristics identical to those of membrane bound receptor. Stability studies indicate that the solubilized cardiac ..beta..AR has a t/sub 1/2/ of 92 hours. Solubilized receptor is stabilized by occupation with antagonists. Isoelectric focusing indicates a pI = 5.0, in agreement with results obtained for both ..beta../sub 1/AR and ..beta../sub 2/AR isolated from other sources. The cardiac ..beta../sub 1/AR has been purified to homogeneity by affinity chromatography and size exclusion HPLC (2 TSK 2000, 1 TSK 3000). Autoradiograms of purified, radioiodinated receptor preparations subjected to SDS-PAGE revealed a single band with an apparent subunit molecular mass (M/sub r/) of 68 kDa. This subunit M/sub r/ was confirmed in membranes photoaffinity labeled with /sup 125/I-iodocyanopindolol diazirine. A single band was specifically labeled, as evidenced by blocking of photoincorporation by (-) and (+) propranolol with typical ..beta..AR stereoselectivity.

• Research Organization: National Institutes of Health, Bethesda, MD
• OSTI ID: 5247190
• Report Number(s): CONF-8606151-
• Journal Information: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:6; ISSN FEPRA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ANIMALS
AUTONOMIC NERVOUS SYSTEM AGENTS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CARDIOVASCULAR SYSTEM
CELL CONSTITUENTS
CELL MEMBRANES
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DOMESTIC ANIMALS
DRUGS
ELECTRON CAPTURE RADIOISOTOPES
HEART
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
KINETICS
LIGANDS
LIQUID COLUMN CHROMATOGRAPHY
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
MOLECULAR STRUCTURE
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PROTEINS
PURIFICATION
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
SEPARATION PROCESSES
SWINE
SYMPATHOMIMETICS
VERTEBRATES