Reconstitution of water channel function of aquaporins 1 and 2 by expression in yeast secretory vesicles

Larry A. Coury, John C. Mathai, G. V Ramesh Prasad, Jeffrey L. Brodsky, Peter C Agre, Mark L. Zeidel

Research output: Contribution to journalArticle

Abstract

Aquaporins 1 (AQP1) and 2 (AQP2) were expressed in the yeast secretory mutant sec6-4. The mutant accumulates post-Golgi, plasma membrane-targeted vesicles and may be used to produce large quantities of membrane proteins. AQP1 or AQP2 were inducibly expressed in yeast and were localized within isolated sec6-4 vesicles by immunoblot analysis. Secretory vesicles containing AQP1 and AQP2 exhibited high water permeabilities and low activation energies for water flow, indicating expression of functional AQP1 and AQP2. AQP1 solubilized from secretory vesicles was successfully reconstituted into proteoliposomes, demonstrating the ability to use the yeast system to express aquaporins for reconstitution studies. The AQP2- containing secretory vesicles showed no increased permeability toward formamide, urea, glycerol, or protons compared with control vesicles, demonstrating that AQP2 is highly selective for water over these other substances. We conclude that the expression of aquaporins in yeast sec6 vesicles is a valid system to further study mammalian water channel function.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Renal Physiology
Volume274
Issue number1 43-1
StatePublished - Jan 1998

Fingerprint

Aquaporin 2
Aquaporin 1
Aquaporins
Secretory Vesicles
Yeasts
Water
Permeability
Glycerol
Urea
Protons
Membrane Proteins
Cell Membrane

Keywords

  • Activation energy
  • Heterologous protein expression
  • Proton permeability
  • Water and solute permeability

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Cite this

Reconstitution of water channel function of aquaporins 1 and 2 by expression in yeast secretory vesicles. / Coury, Larry A.; Mathai, John C.; Prasad, G. V Ramesh; Brodsky, Jeffrey L.; Agre, Peter C; Zeidel, Mark L.

In: American Journal of Physiology - Renal Physiology, Vol. 274, No. 1 43-1, 01.1998.

Research output: Contribution to journalArticle

Coury, Larry A. ; Mathai, John C. ; Prasad, G. V Ramesh ; Brodsky, Jeffrey L. ; Agre, Peter C ; Zeidel, Mark L. / Reconstitution of water channel function of aquaporins 1 and 2 by expression in yeast secretory vesicles. In: American Journal of Physiology - Renal Physiology. 1998 ; Vol. 274, No. 1 43-1.
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