Colocalization and FRET-analysis of subunits c and a of the vacuolar H+-ATPase in living plant cells

Seidel T, Kluge C, Hanitzsch M, Ross J, Sauer M, Dietz K-J, Golldack D (2004)
JOURNAL OF BIOTECHNOLOGY 112(1-2): 165-175.

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Zeitschriftenaufsatz | Veröffentlicht | Englisch
Abstract / Bemerkung
The proton-translocating plant vacuolar H+-ATPase (VHA) is of prime importance for acidification of intracellular compartments and is essential for processes such as secondary activated transport, maintenance of ion homeostasis, and adaptation to environmental stress. Twelve genes have been identified that encode subunits of the functional V-ATPase complex. In this study, subunits c and a of the V-ATPase from the plant Mesembryanthemum crystallinum were fused to cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP), respectively, and were transiently coexpressed in protoplasts. Two-colour scanning confocal fluorescence microscopy demonstrates that the fusion proteins VHA-c-CFP and VHA-a-YFP are colocalized at the tonoplast, the plasmamembrane, and at endoplasmic membrane structures indicating expression in cytoplasmic vesicles. Furthermore, fluorescence resonance energy transfer (FRET) was used to visualize the interaction of VHA-c and VHA-a in vivo on the nanometer length scale. Excitation of CFP as donor fluorophore caused increased emission of YFP-fluorescence in protoplasts due to FRET. Our results give strong evidence for physical interaction of subunits c and a in living plant cells. (C) 2004 Elsevier B.V. All rights reserved.
Erscheinungsjahr
Zeitschriftentitel
JOURNAL OF BIOTECHNOLOGY
Band
112
Zeitschriftennummer
1-2
Seite
165-175
ISSN
PUB-ID

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Seidel T, Kluge C, Hanitzsch M, et al. Colocalization and FRET-analysis of subunits c and a of the vacuolar H+-ATPase in living plant cells. JOURNAL OF BIOTECHNOLOGY. 2004;112(1-2):165-175.
Seidel, T., Kluge, C., Hanitzsch, M., Ross, J., Sauer, M., Dietz, K. - J., & Golldack, D. (2004). Colocalization and FRET-analysis of subunits c and a of the vacuolar H+-ATPase in living plant cells. JOURNAL OF BIOTECHNOLOGY, 112(1-2), 165-175. doi:10.1016/j.jbiotec.2004.04.027
Seidel, T., Kluge, C., Hanitzsch, M., Ross, J., Sauer, M., Dietz, K. - J., and Golldack, D. (2004). Colocalization and FRET-analysis of subunits c and a of the vacuolar H+-ATPase in living plant cells. JOURNAL OF BIOTECHNOLOGY 112, 165-175.
Seidel, T., et al., 2004. Colocalization and FRET-analysis of subunits c and a of the vacuolar H+-ATPase in living plant cells. JOURNAL OF BIOTECHNOLOGY, 112(1-2), p 165-175.
T. Seidel, et al., “Colocalization and FRET-analysis of subunits c and a of the vacuolar H+-ATPase in living plant cells”, JOURNAL OF BIOTECHNOLOGY, vol. 112, 2004, pp. 165-175.
Seidel, T., Kluge, C., Hanitzsch, M., Ross, J., Sauer, M., Dietz, K.-J., Golldack, D.: Colocalization and FRET-analysis of subunits c and a of the vacuolar H+-ATPase in living plant cells. JOURNAL OF BIOTECHNOLOGY. 112, 165-175 (2004).
Seidel, Thorsten, Kluge, C, Hanitzsch, M, Ross, J, Sauer, Markus, Dietz, Karl-Josef, and Golldack, Dortje. “Colocalization and FRET-analysis of subunits c and a of the vacuolar H+-ATPase in living plant cells”. JOURNAL OF BIOTECHNOLOGY 112.1-2 (2004): 165-175.

28 Zitationen in Europe PMC

Daten bereitgestellt von Europe PubMed Central.

Using intrinsically fluorescent proteins for plant cell imaging.
Dixit R, Cyr R, Gilroy S., Plant J 45(4), 2006
PMID: 16441351
Imaging protein molecules using FRET and FLIM microscopy.
Wallrabe H, Periasamy A., Curr Opin Biotechnol 16(1), 2005
PMID: 15722011
Mapping of C-termini of V-ATPase subunits by in vivo-FRET measurements.
Seidel T, Golldack D, Dietz KJ., FEBS Lett 579(20), 2005
PMID: 16061227

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