Influence of bacterial toxins and forskolin upon vasopressin-induced inositol phosphate accumulation in WRK 1 cells

Biochem J. 1989 Jun 15;260(3):665-72. doi: 10.1042/bj2600665.

Abstract

The accumulation of inositol phosphates in WRK 1 cells, stimulated with a range of vasopressin concentrations, was diminished by prior exposure to cholera toxin or forskolin, whilst that observed in the presence of maximal concentrations of the hormone was enhanced in pertussis-toxin-treated cells. In the presence of [32P]NAD+, both cholera toxin and pertussis toxin provoked the labelling of peptides with approximate Mrs of 45,000 and 41,000 respectively in the membranes of WRK 1 cells. Exposure to cholera toxin or forskolin for 15-18 h enhanced cyclic AMP accumulation in these cells. The concentrations of these agents which provoked half-maximal cyclic AMP accumulation were similar to those required to diminish receptor-mediated inositol phosphate accumulation by 50%. In contrast, half-maximal ADP-ribosylation of the 45,000Mr peptide needed 100-fold greater concentrations of the toxin than were effective in provoking half-maximal inhibition of inositol phosphate accumulation. Cholera toxin or forskolin also reduced the maximal specific binding, to intact WRK 1 cells, of both [3H][Arg8]vasopressin and the V1a antagonist [3H][beta-mercapto-beta,beta-cyclopentamethylenepropionic acid,O-methyl-Tyr2, Arg8]vasopressin. The kinetics for the loss of this binding capacity following cholera-toxin treatment were very similar to those describing the diminution of vasopressin-stimulated inositol phosphate accumulation in the same cells.

MeSH terms

  • Adenosine Diphosphate / metabolism
  • Animals
  • Cholera Toxin / pharmacology*
  • Colforsin / pharmacology*
  • Cyclic AMP / metabolism
  • Inositol Phosphates / metabolism
  • Mammary Neoplasms, Experimental / metabolism
  • Pertussis Toxin*
  • Rats
  • Receptors, Angiotensin / drug effects
  • Receptors, Vasopressin
  • Tumor Cells, Cultured / drug effects
  • Tumor Cells, Cultured / metabolism
  • Vasopressins / pharmacology*
  • Virulence Factors, Bordetella / pharmacology*

Substances

  • Inositol Phosphates
  • Receptors, Angiotensin
  • Receptors, Vasopressin
  • Virulence Factors, Bordetella
  • Vasopressins
  • Colforsin
  • Adenosine Diphosphate
  • Cholera Toxin
  • Cyclic AMP
  • Pertussis Toxin