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PMID:12640140
| Citation |
Luo, Y, Vassilev, PM, Li, X, Kawanabe, Y and Zhou, J (2003) Native polycystin 2 functions as a plasma membrane Ca2+-permeable cation channel in renal epithelia. Mol. Cell. Biol. 23:2600-7 |
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| Abstract |
Mutations in polycystin 2 (PC2), a Ca(2+)-permeable cation channel, cause autosomal dominant polycystic kidney disease. Whether PC2 functions in the endoplasmic reticulum (ER) or in the plasma membrane has been controversial. Here we generated and characterized a polyclonal antibody against PC2, determined the subcellular localization of both endogenous and transfected PC2 by immunohistochemistry and biotinylation of cell surface proteins, and assessed PC2 channel properties with electrophysiology. Endogenous PC2 was found in the plasma membrane and the primary cilium of mouse inner medullar collecting duct (IMCD) cells and Madin-Darby canine kidney (MDCK) cells, whereas heterologously expressed PC2 showed a predominant ER localization. Patch-clamping of IMCD cells expressing endogenous or heterologous PC2 confirmed the presence of the channel on the plasma membrane. Treatment with chaperone-like factors facilitated the translocation of the PC2 channel to the plasma membrane from intracellular pools. The unitary conductances, channel kinetics, and other characteristics of both endogenously and heterologously expressed PC2 were similar to those described in our previous study in Xenopus laevis oocytes. These results show that PC2 functions as a plasma membrane channel in renal epithelia and suggest that PC2 contributes to Ca(2+) entry and transport of other cations in defined nephron segments in vivo. |
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| Keywords |
Animals; Antibody Specificity; Biotinylation; Calcium/metabolism; Cations/metabolism; Cell Membrane/metabolism; Cells, Cultured; Dogs; Epithelial Cells/chemistry; Epithelial Cells/metabolism; Fluorescent Antibody Technique; Ion Channels/physiology; Kidney/cytology; Kidney/metabolism; Membrane Proteins/analysis; Membrane Proteins/genetics; Membrane Proteins/metabolism; Mice; Potassium/metabolism; Precipitin Tests; Subcellular Fractions/chemistry; TRPP Cation Channels; Transfection |
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