ArticleBiophysical journal2000
cAMP regulated membrane diffusion of a green fluorescent protein-aquaporin 2 chimera.
Article in Biophysical journal, 2000. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 65 citations in OpenAlex.
- Ion channels and actin: A tale of two friends.Histology and histopathology · 2025Review
- Methods for studying mammalian aquaporin biology.Biology methods & protocols · 2023Review
- A multiscale model of the regulation of aquaporin 2 recycling.NPJ systems biology and applications · 2022Article
- Aquaporin-3 in Cancer.International journal of molecular sciences · 2017Review
- AQP2 Plasma Membrane Diffusion Is Altered by the Degree of AQP2-S256 Phosphorylation.International journal of molecular sciences · 2016Article
- Structural Determinants of Oligomerization of the Aquaporin-4 Channel.The Journal of biological chemistry · 2016Article
- The Trafficking of the Water Channel Aquaporin-2 in Renal Principal Cells-a Potential Target for Pharmacological Intervention in Cardiovascular Diseases.Frontiers in pharmacology · 2016Review
- Article
- Easy measurement of diffusion coefficients of EGFP-tagged plasma membrane proteins using k-Space Image Correlation Spectroscopy.Journal of visualized experiments : JoVE · 2014Article
- Expression of aquaporin-3 in ipsilateral rat kidney with unilateral partial ureteral obstruction.Korean journal of urology · 2013Article
- Membrane protein dynamics and functional implications in mammalian cells.Current topics in membranes · 2013Article
- Protein diffusion in plant cell plasma membranes: the cell-wall corral.Frontiers in plant science · 2013Article
- Cell wall constrains lateral diffusion of plant plasma-membrane proteins.Proceedings of the National Academy of Sciences of the United States of America · 2012Article
- Light inactivation of water transport and protein-protein interactions of aquaporin-Killer Red chimeras.The Journal of general physiology · 2012Article
- Aquaporin-4 (AQP4) associations and array dynamics probed by photobleaching and single-molecule analysis of green fluorescent protein-AQP4 chimeras.The Journal of biological chemistry · 2010Article
- Determinants of aquaporin-4 assembly in orthogonal arrays revealed by live-cell single-molecule fluorescence imaging.Journal of cell science · 2009Article
- Cyclic 3',5'-AMP causes ADAM1/ADAM2 to rapidly diffuse within the plasma membrane of guinea pig sperm.Biology of reproduction · 2008Article
- Actin cytoskeleton-dependent dynamics of the human serotonin1A receptor correlates with receptor signaling.Biophysical journal · 2008Article
- Calcium signaling in vasopressin-induced aquaporin-2 trafficking.Pflugers Archiv : European journal of physiology · 2008Review
- A33 antigen displays persistent surface expression.Cancer immunology, immunotherapy : CII · 2008Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 2 countries.
Funding
Abstract
To study the membrane mobility of aquaporin water channels, clones of stably transfected LLC-PK1 cells were isolated with plasma membrane expression of GFP-AQP1 and GFP-AQP2, in which the green fluorescent protein (GFP) was fused upstream and in-frame to each aquaporin (AQP). The GFP fusion did not affect AQP tetrameric association or water transport function. GFP-AQP lateral mobility was measured by irreversibly bleaching a spot (diameter 0.8 microm) on the membrane with an Argon laser beam (488 nm) and following the fluorescence recovery into the bleached area resulting from GFP translational diffusion. In cells expressing GFP-AQP1, fluorescence recovered to >96% of its initial level with t(1/2) of 38 +/- 2 s (23 degrees C) and 21 +/- 1 s (37 degrees C), giving diffusion coefficients (D) of 5.3 and 9.3 x 10(-11) cm(2)/s. GFP-AQP1 diffusion was abolished by paraformaldehyde fixation, slowed >50-fold by the cholesterol-binding agent filipin, but not affected by cAMP agonists. In cells expressing GFP-AQP2, fluorescence recovered to >98% with D of 5.7 and 9.0 x 10(-11) cm(2)/s at 23 degrees C and 37 degrees C. In contrast to results for GFP-AQP1, the cAMP agonist forskolin slowed GFP-AQP2 mobility by up to tenfold. The cAMP slowing was blocked by actin filament disruption with cytochalasin D, by K(+)-depletion in combination with hypotonic shock, and by mutation of the protein kinase A phosphorylation consensus site (S256A) at the AQP2 C-terminus. These results indicate unregulated diffusion of AQP1 in membranes, but regulated AQP2 diffusion that was dependent on phosphorylation at serine 256, and an intact actin cytoskeleton and clathrin coated pit. The cAMP-induced immobilization of phosphorylated AQP2 provides evidence for AQP2-protein interactions that may be important for retention of AQP2 in specialized membrane domains for efficient membrane recycling.
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Registered trials
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