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Membrane trafficking of AQP5 and cAMP dependent phosphorylation in bronchial epithelium |
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| Authors: | Woo Janghee Chae Young Kwang Jang Se Jin Kim Myoung Sook Baek Jin Hyen Park Jong Chul Trink Barry Ratovitski Edward Lee Taekyul Park Beomsoo Park Minjoo Kang Ji Hye Soria Jean Charles Lee Juna Califano Joseph Sidransky David Moon Chulso |
| Affiliation: | a Department of Otolaryngology, Johns Hopkins University, 1550 Orleans St. 574A, Baltimore, MD 21231, USA b Institute for Cell Engineering, Johns Hopkins University, 1550 Orleans St. 574A, Baltimore, MD 21231, USA c Graduate Program in Human Genetics, Johns Hopkins University, 1550 Orleans St. 574A, Baltimore, MD 21231, USA d Department of Oncology, Johns Hopkins University, 1550 Orleans St. 574A, Baltimore, MD 21231, USA e Department of Pathology, Asan Medical Center, Seoul 138-736, Republic of Korea f Gustave-Roussy Institute, Division of Cancer Medicine, 39 Rue Camille Desmoulins, 94805 Villejuif, France |
| Abstract: | Phosphorylation pathway has been identified as an important step in membrane trafficking for AQP5. We generated stably transfected BEAS-2B human bronchial epithelial cells with various over-expression constructs on permeable support. In stable cells with wild-type AQP5 and S156A (AQP5 mutant targeting PKA consensus sequence), AQP5 expression was predominantly polarized to the apical membrane, whereas stable cells with N185D (AQP5 mutant targeting second NPA motif), mainly localized to the cytoplasm. Treatment with H89 and/or chlorophenylthio-cAMP (cpt-cAMP) did not affect membrane expression of AQP5 in any of three stable cells. In cells with wild-type AQP5 and N185D, AQP5s were phosphorylated by PKA, while phosphorylation of AQP5 was not detected in cells with S156A. These results indicate that, in AQP5, serine156 may be phosphorylated by PKA, but membrane expression of AQP5 may not be regulated by PKA phosphorylation. We conclude that AQP5 membrane targeting can include more than one mechanism besides cAMP dependent phosphorylation. |
| Keywords: | Membrane localization Aquaporin AQP5 cAMP dependent phosphorylation |
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