Solubilization and molecular size of atrial natriuretic hormone (ANH) receptors from rabbit aorta, renal cortex and adrenal

Conference · Thu May 01 04:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5022992

Budzik, G P; Bush, E N; Holleman, W H

ANH(1-28) is presumed to regulate blood pressure and fluid balance via membrane receptors coupled to particulate guanylate cyclase. ANH receptors were solubilized from rabbit aorta, renal cortex and adrenal, primary ANH targets. Plasma membranes extracted with 3-((3-cholamidopropyl)dimethylammonio)-1-propane sulfonate(CHAPS) yield solubilized receptors with high affinity binding of /sup 125/I-Tyr/sup 28/-ANH. Degradation of hormone was minimized with a broad spectrum of protease inhibitors. /sup 125/I-ANH binding reached maximum by 1 hr at 0/sup 0/C and was stable for at least an additional 2 hrs. Bound was separated from free ligand by HPLC gel filtration on TSK-3000SW in PBS/CHAPS. Bound hormone eluted at a MW of approx. 200KD in each tissue preparation and was displaced by unlabelled ANH. The concentration of solubilized binding sites was proportional to densities in intact plasma membranes, i.e., adrenal > renal > aorta. Following separation of free hormone, /sup 125/I-ANH-receptors complexes were coupled using bifunctional crosslinking reagents. SDS-PAGE analysis and autoradiography indicated a major labelled band at approx. 150KD in each tissue preparation. The mobility of this labelled band was not sensitive to reduction before SDS-PAGE. Although these results suggest that solubilized ANH receptors from primary target tissues are very similar, microheterogeneity affecting binding affinity or signal transduction cannot as yet be excluded.

• Research Organization: Abbott Labs., Abbott Park, IL
• OSTI ID: 5022992
• Report Number(s): CONF-8606151-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:6
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ADRENAL GLANDS
AFFINITY
ANIMALS
AORTA
ARTERIES
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BLOOD PRESSURE
BLOOD VESSELS
BODY
CARDIOVASCULAR SYSTEM
CELL CONSTITUENTS
CELL MEMBRANES
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
ENDOCRINE GLANDS
GLANDS
HORMONES
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPES
KIDNEYS
KINETICS
LIQUID COLUMN CHROMATOGRAPHY
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PROTEINS
RABBITS
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
SEPARATION PROCESSES
SIZE
SOLVENT PROPERTIES
VERTEBRATES