Membrane trafficking of AQP5 and cAMP dependent phosphorylation in bronchial epithelium.
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.
Department of Otolaryngology, Johns Hopkins University, 1550 Orleans St. 574A, Baltimore, MD 21231, USA.
Janghee
J
Woo
Young Kwang
YK
Chae
Se Jin
SJ
Jang
Myoung Sook
MS
Kim
Jin Hyen
JH
Baek
Jong Chul
JC
Park
Barry
B
Trink
Edward
E
Ratovitski
Taekyul
T
Lee
Beomsoo
B
Park
Minjoo
M
Park
Ji Hye
JH
Kang
Jean Charles
JC
Soria
Juna
J
Lee
Joseph
J
Califano
David
D
Sidransky
Chulso
C
Moon
D051402:Aquaporin 5 / Q000378:metabolism*; D001980:Bronchi / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism*; D000242:Cyclic AMP / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism*; D006801:Humans; D010766:Phosphorylation; D021381:Protein Transport / Q000502:physiology*
C496036:AQP5 protein, human; D051402:Aquaporin 5; D000242:Cyclic AMP