Regulatory assembly of the vacuolar proton pump VoV1-ATPase in yeast cells by FLIM-FRET

Stefan Ernst; Claire Batisse; Nawid Zarrabi; Bettina Böttcher; Michael Börsch

doi:10.1117/12.841169

26 February 2010 Regulatory assembly of the vacuolar proton pump V_oV₁-ATPase in yeast cells by FLIM-FRET

Stefan Ernst, Claire Batisse, Nawid Zarrabi, Bettina Böttcher, Michael Börsch

Proceedings Volume 7569, Multiphoton Microscopy in the Biomedical Sciences X; 75690W (2010) https://doi.org/10.1117/12.841169
Event: SPIE BiOS, 2010, San Francisco, California, United States

Abstract

We investigate the reversible disassembly of V_OV₁-ATPase in life yeast cells by time resolved confocal FRET imaging. V_OV₁-ATPase in the vacuolar membrane pumps protons from the cytosol into the vacuole. V_OV₁-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 V_OV₁-ATPase is regulated through assembly / disassembly processes. During starvation the two parts of V_OV₁-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.

Citation Download Citation

Stefan Ernst, Claire Batisse, Nawid Zarrabi, Bettina Böttcher, and Michael Börsch "Regulatory assembly of the vacuolar proton pump V_oV₁-ATPase in yeast cells by FLIM-FRET", Proc. SPIE 7569, Multiphoton Microscopy in the Biomedical Sciences X, 75690W (26 February 2010); https://doi.org/10.1117/12.841169

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