Regulatory assembly of the vacuolar proton pump VOV1-ATPase in yeast cells by FLIM-FRET

Details Regulatory assembly of the vacuolar proton pump VOV1-ATPase in yeast cells by FLIM-FRET Regulatory assembly of the vacuolar proton pump VOV1-ATPase in yeast cells by FLIM-FRET

2010-02-02

arxiv.org/abs/1002.0409v1

Biology

Quantitative Biology

Subcellular Processes

8 pages, 3 figures

Scientific paper

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 transport processes as well as for pH and Ca2+ homoeostasis in the cell. 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.

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