Ligand-induced cleavage of the V2 vasopressin receptor by a plasma membrane metalloproteinase.
The proteolytic cleavage of a G protein-coupled peptide hormone receptor, the renal V2 vasopressin receptor, by a plasma membrane proteinase was investigated. In the absence of protease inhibitors during incubation of bovine kidney membranes with a photoreactive vasopressin agonist, V2 receptor truncation leads to a labeled receptor fragment with M(r) 30,000. The V2 receptor-degrading enzyme could be completely inhibited by zinc ions yielding the native V2 receptor glycoprotein with M(r) 58,000. Studies with inhibitors of metalloendopeptidases involved in peptide hormone metabolism and with peptide substrates spanning the V2 receptor cleavage site classify the receptor protease as metalloendoproteinase with specificity for longer substrates. Comparison of the NH2-terminal protein sequence of the truncated M(r) 30,000 V2 receptor with the sequence deduced from the cDNA of the cloned bovine V2 receptor shows that cleavage occurs between Gln92 and Val93 of the second transmembrane helix close to an extracellular agonist binding site. V2 receptor proteolysis was dependent on the presence of a hormonal ligand. It occurred rapidly after hormone binding and led to a loss of ligand binding properties of the truncated V2 receptor. The data suggest that the endogenous V2 receptor-degrading metalloendoproteinase regulates V2 receptor function. The novel pathway may contribute to the termination of signal transmission.
Kojro
E
E
Max-Planck-Institute of Biophysics, Frankfurt am Main, Federal Republic of Germany.
Fahrenholz
F
F
D000595:Amino Acid Sequence; D000818:Animals; D001127:Arginine Vasopressin / Q000378:metabolism*; D002417:Cattle; D006031:Glycosylation; D008024:Ligands; D008666:Metalloendopeptidases / Q000502:physiology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011480:Protease Inhibitors / Q000494:pharmacology; D017483:Receptors, Vasopressin / Q000378:metabolism*; D015032:Zinc / Q000494:pharmacology
D008024:Ligands; D011480:Protease Inhibitors; D017483:Receptors, Vasopressin; D001127:Arginine Vasopressin; D008666:Metalloendopeptidases; D015032:Zinc