Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements

Seidel T, Golldack D, Dietz K-J (2005)
FEBS Letters 579(20): 4374-4382.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
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Abstract / Bemerkung
The plant V-ATPase is a protein complex of 13 different VHA-subunits and functions as ATP driven motor that electrogenically translocates H+ into endomembrane compartments. The central rotor extends into the hexameric head that is fixed by peripheral stators to an eccentric membrane domain. The localization and orientation of VHA-subunits of the head and peripheral stalk region were investigated by in vivo fluorescence resonance energy transfer (FRET). To this end, VHA-E, VHA-G, VHA-H of the peripheral stalks as well as subunits VHA-A and VHA-B were C-terminally fused to cyan (CFP) and yellow fluorescent protein (YFP). Protoplasts transfected with FRET-pairs of CFP-donor and YFP-acceptor fluorophores fused to VHA-subunits were analysed for FRET by laser scanning microscopy. The result of the C-termini mapping allows to refine the arrangement and interaction of the subunits within the V-ATPase complex in vivo. Furthermore, expression of fused VHA-E and VHA-H stimulated acidification of protoplast vacuoles, while other constructs had no major effect on vacuolar pH tentatively indicating a regulatory role of these subunits in plants. (c) 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
FRET; plant; peripheral stalk; V-ATPase
FEBS Letters
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Seidel T, Golldack D, Dietz K-J. Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements. FEBS Letters. 2005;579(20):4374-4382.
Seidel, T., Golldack, D., & Dietz, K. - J. (2005). Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements. FEBS Letters, 579(20), 4374-4382.
Seidel, Thorsten, Golldack, Dortje, and Dietz, Karl-Josef. 2005. “Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements”. FEBS Letters 579 (20): 4374-4382.
Seidel, T., Golldack, D., and Dietz, K. - J. (2005). Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements. FEBS Letters 579, 4374-4382.
Seidel, T., Golldack, D., & Dietz, K.-J., 2005. Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements. FEBS Letters, 579(20), p 4374-4382.
T. Seidel, D. Golldack, and K.-J. Dietz, “Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements”, FEBS Letters, vol. 579, 2005, pp. 4374-4382.
Seidel, T., Golldack, D., Dietz, K.-J.: Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements. FEBS Letters. 579, 4374-4382 (2005).
Seidel, Thorsten, Golldack, Dortje, and Dietz, Karl-Josef. “Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements”. FEBS Letters 579.20 (2005): 4374-4382.

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