Solubilization and reconstitution of renal vasopressin receptors

Conference · Fri May 01 04:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5904366

Aiyar, N; Nambi, P; Stassen, F; Crooke, S T

Renal vasopressin receptors (V/sub 2/) mediating antidiuresis are coupled to adenylate cyclase. To determine the molecular properties of these vasopressin receptors, it is necessary to solubilize the receptors from the membranes. Solubilization of vasopressin receptors in the non-liganded state was shown to abolish hormone recognition. To preserve ligand binding capacity they have developed reconstitution procedures for the renal vasopressin receptors. The pig kidney membranes were solubilized using a zwitterionic detergent, egg lysolecithin and then reconstituted into phospholipid vesicles. Specific binding of (/sup 3/H) lysine vasopressin ((/sup 3/H)LVP) to these solubilized reconstituted fractions was fast, saturable and increased linearly with protein concentration. Scatchard analysis of (/sup 3/H)LVP binding indicated the presence of single class of binding sites with an equilibrium dissociation constant of 2.3 nM. In competition binding experiments, the solubilized receptors displayed the same pharmacological profile as was observed with membrane V/sub 2/ receptors.

• Research Organization: SK and F Labs., Philadelphia, PA
• OSTI ID: 5904366
• Report Number(s): CONF-870644-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 46:6
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
BIOCHEMICAL REACTION KINETICS
BODY
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL PROPERTIES
CYCLASES
DOMESTIC ANIMALS
ENZYMES
HORMONES
ISOTOPE APPLICATIONS
KIDNEYS
KINETICS
LABELLED COMPOUNDS
LYASES
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
ORGANIC COMPOUNDS
ORGANS
PEPTIDE HORMONES
PITUITARY HORMONES
PROTEINS
REACTION KINETICS
RECEPTORS
SOLUBILITY
SWINE
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VASOPRESSIN
VERTEBRATES