AJP - Renal Track the topics, authors and articles important to you
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Am J Physiol Renal Physiol 274: F34-F42, 1998;
0363-6127/98 $5.00
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Coury, L. A.
Right arrow Articles by Zeidel, M. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Coury, L. A.
Right arrow Articles by Zeidel, M. L.
Vol. 274, Issue 1, F34-F42, January 1998

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

Larry A. Coury1, John C. Mathai2, G. V. Ramesh Prasad1, Jeffrey L. Brodsky3, Peter Agre2, and Mark L. Zeidel1

1 Laboratory of Epithelial Cell Biology, Renal Electrolyte Division, Department of Medicine, University of Pittsburgh Medical Center, Pittsburgh 15213-2500; 3 Department of Biological Sciences, University of Pittsburgh, Pittsburgh, Pennsylvania 15260; and 2 Departments of Biological Chemistry and Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205-2185

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.

activation energy; heterologous protein expression; proton permeability; water and solute permeability


This article has been cited by other articles:


Home page
 Am. J. Physiol. Cell Physiol.Home page
N. Kaufmann, J. C. Mathai, W. G. Hill, J. A. T. Dow, M. L. Zeidel, and J. L. Brodsky
Developmental expression and biophysical characterization of a Drosophila melanogaster aquaporin
Am J Physiol Cell Physiol, August 1, 2005; 289(2): C397 - C407.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Cell Physiol.Home page
W. G. Hill, E. Almasri, W. G. Ruiz, G. Apodaca, and M. L. Zeidel
Water and solute permeability of rat lung caveolae: high permeabilities explained by acyl chain unsaturation
Am J Physiol Cell Physiol, July 1, 2005; 289(1): C33 - C41.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Cell Physiol.Home page
V.-K. Ton and R. Rao
Functional expression of heterologous proteins in yeast: insights into Ca2+ signaling and Ca2+-transporting ATPases
Am J Physiol Cell Physiol, September 1, 2004; 287(3): C580 - C589.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
A. S. Verkman and A. K. Mitra
Structure and function of aquaporin water channels
Am J Physiol Renal Physiol, January 1, 2000; 278(1): F13 - F28.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Renal Physiol.Home page
I. Shinbo, K. Fushimi, M. Kasahara, K. Yamauchi, S. Sasaki, and F. Marumo
Functional analysis of aquaporin-2 mutants associated with nephrogenic diabetes insipidus by yeast expression
Am J Physiol Renal Physiol, November 1, 1999; 277(5): F734 - F741.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
L. A. Coury, M. Hiller, J. C. Mathai, E. W. Jones, M. L. Zeidel, and J. L. Brodsky
Water Transport across Yeast Vacuolar and Plasma Membrane-Targeted Secretory Vesicles Occurs by Passive Diffusion
J. Bacteriol., July 15, 1999; 181(14): 4437 - 4440.
[Abstract] [Full Text]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Visit Other APS Journals Online