Aquaporin membrane protein channels mediate cellular water flow. Human aquaporin 5 (AQP5) is highly expressed in the respiratory system and secretory glands where it facilitates the osmotically-driven generation of pulmonary secretions, saliva, sweat and tears. Dysfunctional trafficking of AQP5 has been implicated in several human disease states, including Sjögren’s syndrome, bronchitis and cystic fibrosis. In order to investigate how the plasma membrane expression levels of AQP5 are regulated, we studied real-time translocation of GFP-tagged AQP5 in HEK293 cells. We show that AQP5 plasma membrane abundance in transfected HEK293 cells is rapidly and reversibly regulated by at least three independent mechanisms involving phosphorylation at Ser156, protein kinase A activity and extracellular tonicity. The crystal structure of a Ser156 phosphomimetic mutant indicates that its involvement in regulating AQP5 membrane abundance is not mediated by a conformational change of the carboxy-terminus. We suggest that together these pathways regulate cellular water flow.
Bibliographical note© 2015 Kitchen et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: Swedish Research Council (www.vr.se) (2009-360; 2010-5208); EC FP7 (http://ec.europa.eu/research/fp7/index_en.cfm) (201924 EDICT); and EPSRC (https://www.epsrc.ac.uk) (EP/F500378/1).
Data: Coordinates and structure factors are available from the Protein Data Bank, accession codes 5DYE and 5C5X. All other relevant data are within the paper.
Kitchen, P., Öberg, F., Sjöhamn, J., Hedfalk, K., Bill, R. M., Conner, A. C., Conner, M. T., & Törnroth-Horsefield, S. (2015). Plasma membrane abundance of human aquaporin 5 is dynamically regulated by multiple pathways. PLoS ONE, 10(11), [e0143027]. https://doi.org/10.1371/journal.pone.0143027