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Proceedings Paper

Regulatory assembly of the vacuolar proton pump VoV1-ATPase in yeast cells by FLIM-FRET
Author(s): Stefan Ernst; Claire Batisse; Nawid Zarrabi; Bettina Böttcher; Michael Börsch
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Paper Abstract

We investigate the reversible disassembly of VOV1-ATPase in life yeast cells by time resolved confocal FRET imaging. VOV1-ATPase in the vacuolar membrane pumps protons from the cytosol into the vacuole. VOV1-ATPase is a rotary biological nanomotor driven by ATP hydrolysis. The emerging proton gradient is used for secondary transport processes as well as for pH and Ca2+ homoeostasis in the cell. The activity of the VOV1-ATPase is regulated through assembly / disassembly processes. During starvation the two parts of VOV1-ATPase start to disassemble. This process is reversed after addition of glucose. The exact mechanisms are unknown. To follow the disassembly / reassembly in vivo we tagged two subunits C and E with different fluorescent proteins. Cellular distributions of C and E were monitored using a duty cycle-optimized alternating laser excitation scheme (DCO-ALEX) for time resolved confocal FRET-FLIM measurements.

Paper Details

Date Published: 26 February 2010
PDF: 8 pages
Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 75690W (26 February 2010); doi: 10.1117/12.841169
Show Author Affiliations
Stefan Ernst, Univ. Stuttgart (Germany)
Claire Batisse, EMBL Heidelberg (Germany)
Nawid Zarrabi, Univ. Stuttgart (Germany)
Bettina Böttcher, EMBL Heidelberg (Germany)
Univ. of Edinburgh (United Kingdom)
Michael Börsch, Univ. Stuttgart (Germany)

Published in SPIE Proceedings Vol. 7569:
Multiphoton Microscopy in the Biomedical Sciences X
Ammasi Periasamy; Peter T. C. So; Karsten König, Editor(s)

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