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.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
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.
Erscheinungsjahr
Zeitschriftentitel
FEBS Letters
Band
579
Zeitschriftennummer
20
Seite
4374-4382
ISSN
PUB-ID

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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. doi:10.1016/j.febslet.2005.06.077
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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