PMID- 10644653
OWN - NLM
STAT- MEDLINE
DCOM- 20000331
LR  - 20171213
IS  - 1931-857X (Print)
IS  - 1522-1466 (Linking)
VI  - 278
IP  - 1
DP  - 2000 Jan
TI  - Localization and regulation of PKA-phosphorylated AQP2 in response to
      V(2)-receptor agonist/antagonist treatment.
PG  - F29-42
AB  - Phosphorylation of Ser(256), in a PKA consensus site, in AQP2 (p-AQP2) appears to
      be critically involved in the vasopressin-induced trafficking of AQP2. In the
      present study, affinity-purified antibodies that selectively recognize AQP2
      phosphorylated at Ser(256) were developed. These antibodies were used to
      determine 1) the subcellular localization of p-AQP2 in rat kidney and 2) changes 
      in distribution and/or levels of p-AQP2 in response to
      [desamino-Cys(1),D-Arg(8)]vasopressin (DDAVP) treatment or V(2)-receptor
      blockade. Immunoelectron microscopy revealed that p-AQP2 was localized in both
      the apical plasma membrane and in intracellular vesicles of collecting duct
      principal cells. Treatment of rats with V(2)-receptor antagonist for 30 min
      resulted in almost complete disappearance of p-AQP2 labeling of the apical plasma
      membrane with only marginal labeling of intracellular vesicles remaining.
      Immunoblotting confirmed a marked decrease in p-AQP2 levels. In control
      Brattleboro rats (BB), lacking vasopressin secretion, p-AQP2 labeling was almost 
      exclusively present in intracellular vesicles. Treatment of BB rats with DDAVP
      for 2 h induced a 10-fold increase in p-AQP2 labeling of the apical plasma
      membrane. The overall abundance of p-AQP2, however, was not increased, as
      determined both by immunoelectron microscopy and immunoblotting. Consistent with 
      this, 2 h of DDAVP treatment of normal rats also resulted in unchanged p-AQP2
      levels. Thus the results demonstrate that AQP2 phosphorylated in Ser(256) is
      present in the apical plasma membrane and in intracellular vesicles and that both
      the intracellular distribution/trafficking, as well as the abundance of p-AQP2,
      are regulated via V(2) receptors by altering phosphorylation and/or
      dephosphorylation of Ser(256) in AQP2.
FAU - Christensen, B M
AU  - Christensen BM
AD  - Department of Cell Biology, Institute of Anatomy, University of Aarhus, DK-8000
      Aarhus, Denmark.
FAU - Zelenina, M
AU  - Zelenina M
FAU - Aperia, A
AU  - Aperia A
FAU - Nielsen, S
AU  - Nielsen S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Am J Physiol Renal Physiol
JT  - American journal of physiology. Renal physiology
JID - 100901990
RN  - 0 (Antibodies)
RN  - 0 (Antidiuretic Hormone Receptor Antagonists)
RN  - 0 (Aqp2 protein, rat)
RN  - 0 (Aquaporin 2)
RN  - 0 (Aquaporin 6)
RN  - 0 (Aquaporins)
RN  - 0 (Benzazepines)
RN  - 0 (Receptors, Vasopressin)
RN  - 11000-17-2 (Vasopressins)
RN  - 17OJ42922Y (mozavaptan)
RN  - EC 2.7.11.11 (Cyclic AMP-Dependent Protein Kinases)
RN  - ENR1LLB0FP (Deamino Arginine Vasopressin)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies/immunology
MH  - Antidiuretic Hormone Receptor Antagonists
MH  - Aquaporin 2
MH  - Aquaporin 6
MH  - Aquaporins/immunology/*metabolism
MH  - Benzazepines/pharmacology
MH  - Binding Sites
MH  - Cyclic AMP-Dependent Protein Kinases/*metabolism
MH  - Deamino Arginine Vasopressin/pharmacology
MH  - Immunoblotting
MH  - Immunohistochemistry
MH  - Kidney Tubules/*metabolism
MH  - Microscopy, Immunoelectron
MH  - Molecular Sequence Data
MH  - Phosphorylation
MH  - Rats
MH  - Rats, Brattleboro
MH  - Rats, Wistar
MH  - Receptors, Vasopressin/agonists/*drug effects
MH  - Subcellular Fractions/metabolism
MH  - Vasopressins/deficiency
EDAT- 2000/02/08 00:00
MHDA- 2000/02/08 00:01
CRDT- 2000/02/08 00:00
PHST- 2000/02/08 00:00 [pubmed]
PHST- 2000/02/08 00:01 [medline]
PHST- 2000/02/08 00:00 [entrez]
AID - 10.1152/ajprenal.2000.278.1.F29 [doi]
PST - ppublish
SO  - Am J Physiol Renal Physiol. 2000 Jan;278(1):F29-42. doi:
      10.1152/ajprenal.2000.278.1.F29.