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(Received for publication, March 24, 1997)
From the Second Department of Internal Medicine, School of
Medicine, Tokyo Medical and Dental University, 1-5-45 Yushima,
Bunkyo-ku, Tokyo 113, Japan
The aquaporin-2 (AQP2) vasopressin water channel
is translocated to the apical membrane upon vasopressin stimulation.
Phosphorylation of serine 256 of AQP2 by cAMP-dependent
protein kinase has been shown, but its relation to
vasopressin-regulated translocation has not been elucidated. To address
this question, wild type (WT) AQP2 and a mutant with alanine in place
of serine 256 of AQP2 (S256A) were expressed in LLC-PK1 cells by
electroporation. Measurements by a stopped-flow light-scattering method
revealed that the osmotic water permeability (Pf)
of LLC-PK1 cells transfected with WT was 69.6 ± 6.5 µm/s
(24.8 ± 2.2 µm/s for mock-transfected), and stimulation by 500 µM 8-(4-chlorophenylthio)-cAMP increased the
Pf by 85 ± 12%. When S256A AQP2 was
transfected, the cAMP-dependent increase in the
Pf was only 8 ± 5%. After cAMP stimulation,
the increase in surface expression of AQP2 determined by surface biotin
labeling was 4 ± 10%, significantly less than that for WT
(88 ± 5%). In addition, an in vivo
[32P]orthophosphate labeling assay demonstrated
significant phosphorylation of WT AQP2 and only minimal phosphorylation
of S256A AQP2 in LLC-PK1 cells. Our results indicated that serine 256 of AQP2 is necessary for regulatory exocytosis and that cAMP-responsive
redistribution of AQP2 may be regulated by phosphorylation of AQP2.
Volume 272, Number 23,
Issue of June 6, 1997
pp. 14800-14804
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
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Y. Yamashita, K. Hirai, Y. Katayama, K. Fushimi, S. Sasaki, and F. Marumo Mutations in sixth transmembrane domain of AQP2 inhibit its translocation induced by vasopression Am J Physiol Renal Physiol, March 1, 2000; 278(3): F395 - F405. [Abstract] [Full Text] [PDF]
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 S. C.D. van IJzendoorn and D. Hoekstra Polarized Sphingolipid Transport from the Subapical Compartment Changes during Cell Polarity Development Mol. Biol. Cell, March 1, 2000; 11(3): 1093 - 1101. [Abstract] [Full Text]
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D. Brown Targeting of membrane transporters in renal epithelia: when cell biology meets physiology Am J Physiol Renal Physiol, February 1, 2000; 278(2): F192 - F201. [Abstract] [Full Text] [PDF]
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C. E. Gustafson, T. Katsura, M. McKee, R. Bouley, J. E. Casanova, and D. Brown Recycling of AQP2 occurs through a temperature- and bafilomycin-sensitive trans-Golgi-associated compartment Am J Physiol Renal Physiol, February 1, 2000; 278(2): F317 - F326. [Abstract] [Full Text] [PDF]
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G Valenti, G Procino, M Carmosino, A Frigeri, R Mannucci, I Nicoletti, and M Svelto The phosphatase inhibitor okadaic acid induces AQP2 translocation independently from AQP2 phosphorylation in renal collecting duct cells J. Cell Sci., January 6, 2000; 113(11): 1985 - 1992. [Abstract] [PDF]
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B. M. Christensen, M. Zelenina, A. Aperia, and S. Nielsen Localization and regulation of PKA-phosphorylated AQP2 in response to V2-receptor agonist/antagonist treatment Am J Physiol Renal Physiol, January 1, 2000; 278(1): F29 - F42. [Abstract] [Full Text] [PDF]
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I. Shinbo, K. Fushimi, M. Kasahara, K. Yamauchi, S. Sasaki, and F. Marumo Functional analysis of aquaporin-2 mutants associated with nephrogenic diabetes insipidus by yeast expression Am J Physiol Renal Physiol, November 1, 1999; 277(5): F734 - F741. [Abstract] [Full Text] [PDF]
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A. Rodriguez, I. Martinez, A. Chung, C. H. Berlot, and N. W. Andrews cAMP Regulates Ca2+-dependent Exocytosis of Lysosomes and Lysosome-mediated Cell Invasion by Trypanosomes J. Biol. Chem., June 11, 1999; 274(24): 16754 - 16759. [Abstract] [Full Text] [PDF]
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 T. Ma and A S Verkman Aquaporin water channels in gastrointestinal physiology J. Physiol., June 1, 1999; 517(2): 317 - 326. [Abstract] [Full Text] [PDF]
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S. NIELSEN, T.-H. KWON, B. M. CHRISTENSEN, D. PROMENEUR, J. FRØKI&Aelig;R, and D. MARPLES Physiology and Pathophysiology of Renal Aquaporins J. Am. Soc. Nephrol., March 1, 1999; 10(3): 647 - 663. [Abstract] [Full Text]
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E. Klussmann, K. Maric, B. Wiesner, M. Beyermann, and W. Rosenthal Protein Kinase A Anchoring Proteins Are Required for Vasopressin-mediated Translocation of Aquaporin-2 into Cell Membranes of Renal Principal Cells J. Biol. Chem., February 19, 1999; 274(8): 4934 - 4938. [Abstract] [Full Text] [PDF]
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J. Frokiaer, D. Marples, H. Valtin, J. F. Morris, M. A. Knepper, and S. Nielsen Low aquaporin-2 levels in polyuric DI +/+ severe mice with constitutively high cAMP-phosphodiesterase activity Am J Physiol Renal Physiol, February 1, 1999; 276(2): F179 - F190. [Abstract] [Full Text] [PDF]
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G. Nishimoto, M. Zelenina, D. Li, M. Yasui, A. Aperia, S. Nielsen, and A. C. Nairn Arginine vasopressin stimulates phosphorylation of aquaporin-2 in rat renal tissue Am J Physiol Renal Physiol, February 1, 1999; 276(2): F254 - F259. [Abstract] [Full Text] [PDF]
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T.-H. Kwon, J. Frokiaer, M. A. Knepper, and S. Nielsen Reduced AQP1, -2, and -3 levels in kidneys of rats with CRF induced by surgical reduction in renal mass Am J Physiol Renal Physiol, November 1, 1998; 275(5): F724 - F741. [Abstract] [Full Text] [PDF]
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K. Maric, A. Oksche, and W. Rosenthal Aquaporin-2 expression in primary cultured rat inner medullary collecting duct cells Am J Physiol Renal Physiol, November 1, 1998; 275(5): F796 - F801. [Abstract] [Full Text] [PDF]
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F. F. Bolander Jr. Transduction pathways involved in rapid hormone receptor regulation in the mammary epithelium Am J Physiol Endocrinol Metab, October 1, 1998; 275(4): E553 - E557. [Abstract] [Full Text] [PDF]
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D. Brown, T. Katsura, and C. E. Gustafson Cellular mechanisms of aquaporin trafficking Am J Physiol Renal Physiol, September 1, 1998; 275(3): F328 - F331. [Abstract] [Full Text] [PDF]
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G. G. MacGregor, J. Z. Xu, C. M. McNicholas, G. Giebisch, and S. C. Hebert Partially active channels produced by PKA site mutation of the cloned renal K+ channel, ROMK2 (kir1.2) Am J Physiol Renal Physiol, September 1, 1998; 275(3): F415 - F422. [Abstract] [Full Text] [PDF]
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G. Valenti, G. Procino, U. Liebenhoff, A. Frigeri, P. A. Benedetti, G. Ahnert-Hilger, B. Nurnberg, M. Svelto, and W. Rosenthal A Heterotrimeric G Protein of the Gi Family Is Required for cAMP-triggered Trafficking of Aquaporin 2 in Kidney Epithelial Cells J. Biol. Chem., August 28, 1998; 273(35): 22627 - 22634. [Abstract] [Full Text] [PDF]
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 T. Ma, B. Yang, M. A. Matthay, and A. S. Verkman Evidence against a Role of Mouse, Rat, and Two Cloned Human T1alpha Isoforms as a Water Channel or a Regulator of Aquaporin-type Water Channels Am. J. Respir. Cell Mol. Biol., July 1, 1998; 19(1): 143 - 149. [Abstract] [Full Text]
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I. Johansson, M. Karlsson, V. K. Shukla, M. J. Chrispeels, C. Larsson, and P. Kjellbom Water Transport Activity of the Plasma Membrane Aquaporin PM28A Is Regulated by Phosphorylation PLANT CELL, March 1, 1998; 10(3): 451 - 460. [Abstract] [Full Text] [PDF]
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H. Kipp, N. Pichetshote, and I. M. Arias Transporters on Demand. INTRAHEPATIC POOLS OF CANALICULAR ATP BINDING CASSETTE TRANSPORTERS IN RAT LIVER J. Biol. Chem., March 2, 2001; 276(10): 7218 - 7224. [Abstract] [Full Text] [PDF]
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M. H. Levin, P. M. Haggie, L. Vetrivel, and A. S. Verkman Diffusion in the Endoplasmic Reticulum of an Aquaporin-2 Mutant Causing Human Nephrogenic Diabetes Insipidus J. Biol. Chem., June 8, 2001; 276(24): 21331 - 21336. [Abstract] [Full Text] [PDF]
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P. M. T. Deen, B. W. M. Van Balkom, P. J. M. Savelkoul, E.-J. Kamsteeg, M. Van Raak, M. L. Jennings, T. R. Muth, V. Rajendran, and M. J. Caplan Aquaporin-2: COOH terminus is necessary but not sufficient for routing to the apical membrane Am J Physiol Renal Physiol, February 1, 2002; 282(2): F330 - F340. [Abstract] [Full Text] [PDF]
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