Isolation and characterization of the rat liver AVP receptor using [125I][d(CH2)5'sarcosine7]AVP

Clin Exp Pharmacol Physiol. 1992 Apr;19(4):253-60. doi: 10.1111/j.1440-1681.1992.tb00447.x.

Abstract

1. A vasopressin (AVP) binding protein was purified from rat liver membranes by an improved method using [125I][d(CH2)5'Sarcosine7]AVP, a selective V1 AVP radioligand and a combination of CHAPS solubilization, gel filtration, lectin affinity and FPLC ion exchange chromatography. 2. The purified protein exhibited a maximum binding activity of 2480 pmol/mg protein with a KD of 4.5 nmol/L, which corresponds to a purification of approximately 26,700-fold. The molecular weight of this protein was 70,000 Da. 3. The binding of [125I][d(CH2)5'Sarcosine7]AVP to the solubilized membranes was dependent on the protein concentration, and was inhibited by the unlabelled peptides [d(CH2)5'Sarcosine7]AVP, AVP, and to a lesser degree by peptides with high V2 receptor affinity, such as 1-desamino-D-AVP and [d(CH2)5'D-Ileu2-Ileu4]AVP. 4. In addition, an AVP anti-idiotypic monoclonal antibody bound to both the partially purified and purified lectin affinity AVP binding protein in a concentration-dependent manner. These results indicate that the purified protein displays similar characteristics to the liver membrane-bound AVP V1 receptor.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal
  • Arginine Vasopressin / analogs & derivatives*
  • Arginine Vasopressin / metabolism*
  • Chromatography, High Pressure Liquid
  • Electrophoresis, Polyacrylamide Gel
  • In Vitro Techniques
  • Kinetics
  • Liver / chemistry*
  • Mice
  • Mice, Inbred BALB C
  • Molecular Weight
  • Phospholipids / metabolism
  • Protein Binding
  • Rats
  • Receptors, Angiotensin / drug effects
  • Receptors, Angiotensin / isolation & purification*
  • Receptors, Vasopressin*

Substances

  • Antibodies, Monoclonal
  • Phospholipids
  • Receptors, Angiotensin
  • Receptors, Vasopressin
  • argipressin, (1-beta-mercapto-beta,beta-cyclopentamethylenepropionic acid)-Sar(7)-
  • Arginine Vasopressin