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Am J Physiol Renal Physiol
2009 Mar 01;2963:F649-57. doi: 10.1152/ajprenal.90682.2008.
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Role of multiple phosphorylation sites in the COOH-terminal tail of aquaporin-2 for water transport: evidence against channel gating.
Moeller HB
,
MacAulay N
,
Knepper MA
,
Fenton RA
.
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Arginine vasopressin (AVP)-regulated phosphorylation of the water channel aquaporin-2 (AQP2) at serine 256 (S256) is essential for its accumulation in the apical plasma membrane of collecting duct principal cells. In this study, we examined the role of additional AVP-regulated phosphorylation sites in the COOH-terminal tail of AQP2 on protein function. When expressed in Xenopus laevis oocytes, prevention of AQP2 phosphorylation at S256A (S256A-AQP2) reduced osmotic water permeability threefold compared with wild-type (WT) AQP2-injected oocytes. In contrast, prevention of AQP2 single phosphorylation at S261 (S261A), S264 (S264A), and S269 (S269A), or all three sites in combination had no significant effect on water permeability. Similarly, oocytes expressing S264D-AQP2 and S269D-AQP2, mimicking AQP2 phosphorylated at these residues, had similar water permeabilities to WT-AQP2-expressing oocytes. The use of high-resolution confocal laser-scanning microscopy, as well as biochemical analysis demonstrated that all AQP2 mutants, with the exception of S256A-AQP2, had equal abundance in the oocyteplasma membrane. Correlation of osmotic water permeability relative to plasma membrane abundance demonstrated that lack of phosphorylation at S256, S261, S264, or S269 had no effect on AQP2 unit water transport. Similarly, no effect on AQP2 unit water transport was observed for the 264D and 269D forms, indicating that phosphorylation of the COOH-terminal tail of AQP2 is not involved in gating of the channel. The use of phosphospecific antibodies demonstrated that AQP2 S256 phosphorylation is not dependent on any of the other phosphorylation sites, whereas S264 and S269 phosphorylation depend on prior phosphorylation of S256. In contrast, AQP2 S261 phosphorylation is independent of the phosphorylation status of S256.
Brown,
Phosphorylation events and the modulation of aquaporin 2 cell surface expression.
2008, Pubmed
Brown,
Phosphorylation events and the modulation of aquaporin 2 cell surface expression.
2008,
Pubmed
Christensen,
Axial heterogeneity in basolateral AQP2 localization in rat kidney: effect of vasopressin.
2003,
Pubmed
Cohen,
The regulation of protein function by multisite phosphorylation--a 25 year update.
2000,
Pubmed
Fenton,
Recent discoveries in vasopressin-regulated aquaporin-2 trafficking.
2008,
Pubmed
Fenton,
Acute regulation of aquaporin-2 phosphorylation at Ser-264 by vasopressin.
2008,
Pubmed
Fushimi,
Phosphorylation of serine 256 is required for cAMP-dependent regulatory exocytosis of the aquaporin-2 water channel.
1997,
Pubmed
Hedfalk,
Aquaporin gating.
2006,
Pubmed
Hoffert,
Quantitative phosphoproteomics of vasopressin-sensitive renal cells: regulation of aquaporin-2 phosphorylation at two sites.
2006,
Pubmed
Hoffert,
Vasopressin-stimulated increase in phosphorylation at Ser269 potentiates plasma membrane retention of aquaporin-2.
2008,
Pubmed
Hoffert,
Dynamics of aquaporin-2 serine-261 phosphorylation in response to short-term vasopressin treatment in collecting duct.
2007,
Pubmed
Jespersen,
Dual-function vector for protein expression in both mammalian cells and Xenopus laevis oocytes.
2002,
Pubmed
,
Xenbase
Johansson,
Water transport activity of the plasma membrane aquaporin PM28A is regulated by phosphorylation.
1998,
Pubmed
,
Xenbase
Kamsteeg,
The subcellular localization of an aquaporin-2 tetramer depends on the stoichiometry of phosphorylated and nonphosphorylated monomers.
2000,
Pubmed
,
Xenbase
Kamsteeg,
An impaired routing of wild-type aquaporin-2 after tetramerization with an aquaporin-2 mutant explains dominant nephrogenic diabetes insipidus.
1999,
Pubmed
,
Xenbase
Katsura,
Protein kinase A phosphorylation is involved in regulated exocytosis of aquaporin-2 in transfected LLC-PK1 cells.
1997,
Pubmed
Kuwahara,
cAMP-dependent phosphorylation stimulates water permeability of aquaporin-collecting duct water channel protein expressed in Xenopus oocytes.
1995,
Pubmed
,
Xenbase
Lande,
Phosphorylation of aquaporin-2 does not alter the membrane water permeability of rat papillary water channel-containing vesicles.
1996,
Pubmed
Leduc-Nadeau,
Elaboration of a novel technique for purification of plasma membranes from Xenopus laevis oocytes.
2007,
Pubmed
,
Xenbase
Levitan,
Modulation of ion channels by protein phosphorylation and dephosphorylation.
1994,
Pubmed
Lu,
The phosphorylation state of serine 256 is dominant over that of serine 261 in the regulation of AQP2 trafficking in renal epithelial cells.
2008,
Pubmed
Marr,
Functionality of aquaporin-2 missense mutants in recessive nephrogenic diabetes insipidus.
2001,
Pubmed
,
Xenbase
McDill,
Congenital progressive hydronephrosis (cph) is caused by an S256L mutation in aquaporin-2 that affects its phosphorylation and apical membrane accumulation.
2006,
Pubmed
Moeller,
Serine 269 phosphorylated aquaporin-2 is targeted to the apical membrane of collecting duct principal cells.
2009,
Pubmed
Mulders,
An aquaporin-2 water channel mutant which causes autosomal dominant nephrogenic diabetes insipidus is retained in the Golgi complex.
1998,
Pubmed
,
Xenbase
Nishimoto,
Arginine vasopressin stimulates phosphorylation of aquaporin-2 in rat renal tissue.
1999,
Pubmed
Nunes,
A fluorimetry-based ssYFP secretion assay to monitor vasopressin-induced exocytosis in LLC-PK1 cells expressing aquaporin-2.
2008,
Pubmed
Park,
Potassium channel phosphorylation in excitable cells: providing dynamic functional variability to a diverse family of ion channels.
2008,
Pubmed
Schulz,
Mechanisms of voltage-gated ion channel regulation: from gene expression to localization.
2008,
Pubmed
Törnroth-Horsefield,
Structural mechanism of plant aquaporin gating.
2006,
Pubmed
van Balkom,
The role of putative phosphorylation sites in the targeting and shuttling of the aquaporin-2 water channel.
2002,
Pubmed
Yao,
Regulation of TRP channels by phosphorylation.
2005,
Pubmed
Zampighi,
A method for determining the unitary functional capacity of cloned channels and transporters expressed in Xenopus laevis oocytes.
1995,
Pubmed
,
Xenbase
Zeuthen,
Water transport by Na+-coupled cotransporters of glucose (SGLT1) and of iodide (NIS). The dependence of substrate size studied at high resolution.
2006,
Pubmed
,
Xenbase
