Reversible inactivation of vasopressin and angiotensin II binding to hepatocyte membranes by a calcium-dependent, cytosolic protein.
A cytosolic protein is described which inhibits the binding of vasopressin and angiotensin to their rat liver receptors in the presence of calcium. The binding of insulin and transferrin was unaffected. Inhibition was temperature-dependent; it was maximal in 10 min at 37 degrees C, but required longer incubation times at lower temperatures. The pH optimum was 7.4. Inhibition also required the presence of calcium, with half-maximal inhibition at 6-8 microM calcium, but did not require any other low molecular weight cofactors. Inhibition could be reversed by washing the membranes at pH 5.5, but not by incubation with EGTA. Sephacryl S-300 chromatography showed that activity eluted in two peaks with approximate molecular weights of 70,000 and 150,000. In the presence of calcium, the inhibitory activity eluted at 150,000; in the absence of calcium, most of the inhibitory activity eluted at 70,000. A radiolabeled cytosolic protein with a molecular weight of 70,000 was eluted from inhibited rat liver membranes at pH 5.5 as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We propose that vasopressin and angiotensin II, which both mobilize calcium in hepatocytes via phosphatidylinositol turnover, can, by this same mechanism, activate a protein(s) which reduces further binding to their receptors.
D000804:Angiotensin II / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism; D011506:Proteins / Q000502:physiology*; D051381:Rats; D011919:Rats, Inbred Strains; D011945:Receptors, Angiotensin / Q000187:drug effects / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017483:Receptors, Vasopressin; D013816:Thermodynamics; D014667:Vasopressins / Q000378:metabolism*
D011506:Proteins; D011945:Receptors, Angiotensin; D011956:Receptors, Cell Surface; D017483:Receptors, Vasopressin; D014667:Vasopressins; D000804:Angiotensin II; D002118:Calcium