Determinants of AQP6 trafficking to intracellular sites versus the plasma membrane in transfected mammalian cells

Eric Beitz, Kun Liu, Masahiro Ikeda, William B. Guggino, Peter Agre, Masato Yasui

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Background information. Most AQPs (aquaporins) function at the plasma membrane, however AQP6 is exclusively localized to membranes of intracellular vesicles in acid-secreting type-A intercalated cells of renal collecting ducts. The intracellular distribution indicates that AQP6 has a function distinct from trans-epithelial water movement. Results. We show by mutational analyses and immunofluorescence that the N-terminus of AQP6 is a determinant for its intracellular localization. Presence or absence at the plasma membrane of AQP6 constructs was confirmed by electrophysiological methods. Addition of a GFP (green fluorescent protein) or a HA (haemagglutinin) epitope tag (GFP-AQP6 or HA-AQP6) to the N-terminus of AQP6, directed AQP6 to the plasma membranes of transfected Madin-Darby canine Kidney cells. In contrast, addition of a GFP tag to the C-terminus (AQP6-GFP) caused the protein to remain intracellular, similar to untagged wild-type AQP6. Replacement of the N-terminus of AQP6 by that of AQP1 also directed AQP6 to the plasma membranes, whereas the N-terminus of AQP6 retained AQP1 in cytosolic sites. Conclusion. Our results suggest that the N-terminus of AQP6 is critical for trafficking of the protein to the intracellular sites. Moreover, our studies provide an approach for future identification of proteins involved in vesicle sorting in the acid-secreting type-A intercalated cells.

Original languageEnglish
Pages (from-to)101-109
Number of pages9
JournalBiology of the Cell
Volume98
Issue number2
DOIs
Publication statusPublished - 2006 Feb
Externally publishedYes

Fingerprint

Green Fluorescent Proteins
Cell Membrane
Hemagglutinins
Water Movements
Intracellular Membranes
Aquaporins
Madin Darby Canine Kidney Cells
Acids
Protein Transport
Fluorescent Antibody Technique
Epitopes
Proteins
Kidney

Keywords

  • Aquaporin
  • H-ATPase
  • Haemagglutinin (HA)
  • Intracellular vesicle
  • Sorting signal

ASJC Scopus subject areas

  • Cell Biology

Cite this

Determinants of AQP6 trafficking to intracellular sites versus the plasma membrane in transfected mammalian cells. / Beitz, Eric; Liu, Kun; Ikeda, Masahiro; Guggino, William B.; Agre, Peter; Yasui, Masato.

In: Biology of the Cell, Vol. 98, No. 2, 02.2006, p. 101-109.

Research output: Contribution to journalArticle

Beitz, Eric ; Liu, Kun ; Ikeda, Masahiro ; Guggino, William B. ; Agre, Peter ; Yasui, Masato. / Determinants of AQP6 trafficking to intracellular sites versus the plasma membrane in transfected mammalian cells. In: Biology of the Cell. 2006 ; Vol. 98, No. 2. pp. 101-109.
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