VIP receptors from porcine liver: High yield solubilization in a GTP-insensitive form

Journal Article · Sun Dec 31 23:00:00 EST 1989 · Life Sciences; (USA)

OSTI ID:5961972

Voisin, T; Couvineau, A; Guijarro, L; Laburthe, M ^[1]

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1. Institut National de la Sante et de la Recherche Medicale, Villejuif (France)

Vasoactive intestinal peptide (VIP) receptors were solubilized from porcine liver membranes using CHAPS. The binding of {sup 125}I-VIP to solubilized receptors was reversible, saturable and specific. Scatchard analysis indicated the presence of one binding site with a Kd of 6.5 {plus minus} 0.3 nM and a Bmax of 1.20 {plus minus} 0.15 pmol/mg protein. Solubilized and membrane-bound receptors displayed the same pharmacological profile since VIP and VIP-related peptides inhibited {sup 125}I-VIP binding to both receptor preparations with the same rank order of potency e.g. VIP>helodermin>rat GRF>rat PHI>secretin>human GRF. GTP inhibited {sup 125}I-VIP binding to membrane-bound receptors but not to solubilized receptors supporting functional uncoupling of VIP receptor and G protein during solubilization. Affinity labeling of solubilized and membrane-bound VIP receptors with {sup 125}I-VIP revealed the presence of a single molecular component with Mr 55,000 in both cases. It is concluded that VIP receptors from porcine liver can be solubilized with a good yield, in a GTP-insensitive, G protein-free form. This represents a major advance towards the purification of VIP receptors.

• OSTI ID: 5961972
• Journal Information: Life Sciences; (USA), Journal Name: Life Sciences; (USA) Vol. 48:2; ISSN 0024-3205; ISSN LIFSA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AFFINITY
ANIMALS
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CELL CONSTITUENTS
CELL MEMBRANES
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
DOMESTIC ANIMALS
ELECTRON CAPTURE RADIOISOTOPES
FRACTIONATION
GLANDS
INHIBITION
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLING
LIVER
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
NUCLEI
NUCLEOTIDES
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PEPTIDES
PROTEINS
RADIOISOTOPES
REACTION KINETICS
RECEPTORS
SEPARATION PROCESSES
SOLUBILITY
SWINE
TRACER TECHNIQUES
VERTEBRATES