Vasoactive intestinal peptide receptor on liver plasma membranes: Solubilization and cross-linking

Journal Article · Thu Nov 01 04:00:00 EST 1990 · Peptides (Fayetteville, New York); (USA)

DOI:https://doi.org/10.1016/0196-9781(90)90160-7· OSTI ID:5684712

Nguyen, T D; Kaiser, L M ^[1]

---

1. Duke Univ., Durham, NC (USA)

The hepatic receptor for VIP was solubilized from rat liver plasma membranes with 1.4% digitonin and shown to conserve its ability to bind to the ligand. This solubilized receptor demonstrated the high affinity and specificity for VIP (KD approximately 1 nM, binding preference: VIP greater than PHI greater than secretin greater than thymosin alpha 1) which were observed with the nonsolubilized VIP receptor on intact liver plasma membranes. {sup 125}I-VIP was next cross-linked to either the solubilized or nonsolubilized receptor using disuccinimido suberate or disuccinimido dithiobis(propionate), and the resulting complexes analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by autoradiography. A broad autoradiographic band which demonstrated a high affinity for VIP was identified at Mr 56,000 (53,000 in the absence of the reducing agent dithiothreitol) for both the solubilized and nonsolubilized receptors. We have thus been able to solubilize from rat liver plasma membranes a receptor with high affinity and specificity for VIP, and confirmed its structural similarity with the native VIP receptor in nonsolubilized membranes using cross-linking techniques.

• OSTI ID: 5684712
• Journal Information: Peptides (Fayetteville, New York); (USA), Journal Name: Peptides (Fayetteville, New York); (USA) Vol. 11:6; ISSN 0196-9781; ISSN PPTDD
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMALS
AUTORADIOGRAPHY
BETA DECAY RADIOISOTOPES
BIOCHEMICAL REACTION KINETICS
BODY
CELL CONSTITUENTS
CELL MEMBRANES
CHEMICAL REACTIONS
CROSS-LINKING
DAYS LIVING RADIOISOTOPES
DIGESTIVE SYSTEM
ELECTRON CAPTURE RADIOISOTOPES
ELECTROPHORESIS
GASTROINTESTINAL TRACT
GLANDS
INTERMEDIATE MASS NUCLEI
INTESTINES
IODINE 125
IODINE ISOTOPES
ISOTOPES
KINETICS
LIGANDS
LIVER
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
MOLECULAR WEIGHT
NUCLEI
ODD-EVEN NUCLEI
ORGANIC COMPOUNDS
ORGANS
PEPTIDES
POLYMERIZATION
PROTEINS
RADIOISOTOPES
RATS
REACTION KINETICS
REAGENTS
RECEPTORS
RODENTS
SOLUBILITY
VERTEBRATES