AQP2 exocytosis in the renal collecting duct - Involvement of SNARE isoforms and the regulatory role of Munc18b

Giuseppe Procino, Claudia Barbieri, Grazia Tamma, Leonarda De Benedictis, Jeffrey E. Pessin, Maria Svelto, Giovanna Valenti

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Vasopressin regulates the fusion of the water channel aquaporin 2 (AQP2) to the apical membrane of the renal collecting-duct principal cells and several lines of evidence indicate that SNARE proteins mediate this process. In this work MCD4 renal cells were used to investigate the functional role of a set of Q-and R-SNAREs, together with that of Munc18b as a negative regulator of the formation of the SNARE complex. Both VAMP2 and VAMP3 were associated with immunoisolated AQP2 vesicles, whereas syntaxin 3 (Stx3), SNAP23 and Munc18 were associated with the apical plasma membrane. Co-immunoprecipitation experiments indicated that Stx3 forms complexes with VAMP2, VAMP3, SNAP23 and Munc18b. Protein knockdown coupled to apical surface biotinylation demonstrated that reduced levels of tht R-SNAREs VAMP2 and VAMP3, and the Q-SNAREs Stx3 and SNAP23 strongly inhibited AQP2 fusion at the apical membrane. In addition, knockdown of Munc18b promoted a sevenfold increase of AQP2 fused at the plasma membrane without forskolin stimulation. Taken together these findings propose VAMP2, VAMP3, Stx3 and SNAP23 as the complementary set of SNAREs responsible for AQP2-vesicle fusion into the apical membrane, and Munc18b as a negative regulator of SNARE-complex formation in renal collecting-duct principal cells.

Original languageEnglish (US)
Pages (from-to)2097-2106
Number of pages10
JournalJournal of Cell Science
Volume121
Issue number12
DOIs
StatePublished - Jun 15 2008

Fingerprint

Aquaporin 2
Vesicle-Associated Membrane Protein 3
SNARE Proteins
Vesicle-Associated Membrane Protein 2
Qa-SNARE Proteins
Exocytosis
Protein Isoforms
Q-SNARE Proteins
Kidney
Membranes
Cell Membrane
Biotinylation
Aquaporins
Colforsin
Vasopressins
Immunoprecipitation
Cell Line
Proteins

Keywords

  • Aquaporin2
  • Exocytosis
  • Munc18
  • SNARE
  • Syntaxin
  • VAMP
  • Vasopressin

ASJC Scopus subject areas

  • Cell Biology

Cite this

AQP2 exocytosis in the renal collecting duct - Involvement of SNARE isoforms and the regulatory role of Munc18b. / Procino, Giuseppe; Barbieri, Claudia; Tamma, Grazia; De Benedictis, Leonarda; Pessin, Jeffrey E.; Svelto, Maria; Valenti, Giovanna.

In: Journal of Cell Science, Vol. 121, No. 12, 15.06.2008, p. 2097-2106.

Research output: Contribution to journalArticle

Procino, G, Barbieri, C, Tamma, G, De Benedictis, L, Pessin, JE, Svelto, M & Valenti, G 2008, 'AQP2 exocytosis in the renal collecting duct - Involvement of SNARE isoforms and the regulatory role of Munc18b', Journal of Cell Science, vol. 121, no. 12, pp. 2097-2106. https://doi.org/10.1242/jcs.022210
Procino, Giuseppe ; Barbieri, Claudia ; Tamma, Grazia ; De Benedictis, Leonarda ; Pessin, Jeffrey E. ; Svelto, Maria ; Valenti, Giovanna. / AQP2 exocytosis in the renal collecting duct - Involvement of SNARE isoforms and the regulatory role of Munc18b. In: Journal of Cell Science. 2008 ; Vol. 121, No. 12. pp. 2097-2106.
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