Arginine vasopressin stimulates phosphorylation of aquaporin-2 in rat renal tissue

Am J Physiol. 1999 Feb;276(2):F254-9. doi: 10.1152/ajprenal.1999.276.2.F254.


Aquaporin-2 (AQP2), the protein that mediates arginine vasopressin (AVP)-regulated apical water transport in the renal collecting duct, possesses a single consensus phosphorylation site for cAMP-dependent protein kinase A (PKA) at Ser256. The aim of this study was to examine whether AVP, and other agents that increase cAMP levels, could stimulate the phosphorylation of AQP2 in intact rat renal tissue. Rat renal papillae were prelabeled with 32P and incubated with vehicle or drugs, and then AQP2 was immunoprecipitated. Two polypeptides corresponding to nonglycosylated (29 kDa) and glycosylated (35-48 kDa) AQP2 were identified by SDS-PAGE. AVP caused a time- and dose-dependent increase in phosphorylation of both glycosylated and nonglycosylated AQP2. The threshold dose for a significant increase in phosphorylation was 10 pM, which corresponds to a physiological serum concentration of AVP. Maximal phosphorylation was reached within 1 min of AVP incubation. This effect on AQP2 phosphorylation was mimicked by the vasopressin (V2) agonist, 1-desamino-[8-D-arginine]vasopressin (DDAVP), or forskolin. Two-dimensional phosphopeptide mapping indicated that AVP and forskolin stimulated the phosphorylation of the same site in AQP2. Immunoblot analysis using a phosphorylation state-specific antiserum revealed an increase in phosphorylation of Ser256 after incubation of papillae with AVP. The results indicate that AVP stimulates phosphorylation of AQP2 at Ser256 via activation of PKA, supporting the idea that this is one of the first steps leading to increased water permeability in collecting duct cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Aquaporin 2
  • Aquaporin 6
  • Aquaporins / chemistry
  • Aquaporins / metabolism*
  • Arginine Vasopressin / pharmacology*
  • Colforsin / pharmacology
  • Deamino Arginine Vasopressin / pharmacology
  • Differential Threshold / physiology
  • Dose-Response Relationship, Drug
  • Glycosylation
  • In Vitro Techniques
  • Kidney Medulla / metabolism*
  • Male
  • Peptide Mapping
  • Phosphorylation / drug effects
  • Rats
  • Rats, Sprague-Dawley
  • Renal Agents / pharmacology
  • Time Factors


  • Aqp2 protein, rat
  • Aquaporin 2
  • Aquaporin 6
  • Aquaporins
  • Renal Agents
  • Arginine Vasopressin
  • Colforsin
  • Deamino Arginine Vasopressin