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

Stefan Ernst; Claire Batisse; Nawid Zarrabi; Bettina Boettcher; Michael Boersch

doi:10.1117/12.841169

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

Stefan Ernst, Claire Batisse, Nawid Zarrabi, Bettina Boettcher, Michael Boersch

School of Biological Sciences

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

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.

Original language	English
Title of host publication	MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES X
Editors	A Periasamy, PTC So, K Konig
Place of Publication	BELLINGHAM
Publisher	SPIE
Pages	-
Number of pages	8
ISBN (Print)	978-0-8194-7965-5
DOIs	https://doi.org/10.1117/12.841169
Publication status	Published - 2010
Event	Conference on Multiphoton Microscopy in the Biomedical Sciences X - San Francisco Duration: 24 Jan 2010 → 26 Jan 2010

Conference

Conference	Conference on Multiphoton Microscopy in the Biomedical Sciences X
City	San Francisco
Period	24/01/10 → 26/01/10

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10.1117/12.841169

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

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.",

author = "Stefan Ernst and Claire Batisse and Nawid Zarrabi and Bettina Boettcher and Michael Boersch",

year = "2010",

doi = "10.1117/12.841169",

language = "English",

isbn = "978-0-8194-7965-5",

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editor = "A Periasamy and PTC So and K Konig",

booktitle = "MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES X",

publisher = "SPIE",

address = "United States",

note = "Conference on Multiphoton Microscopy in the Biomedical Sciences X ; Conference date: 24-01-2010 Through 26-01-2010",

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Ernst, S, Batisse, C, Zarrabi, N, Boettcher, B & Boersch, M 2010, Regulatory assembly of the vacuolar proton pump VOV1-ATPase in yeast cells by FLIM-FRET. in A Periasamy, PTC So & K Konig (eds), MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES X. SPIE, BELLINGHAM, pp. -, Conference on Multiphoton Microscopy in the Biomedical Sciences X, San Francisco, 24/01/10. https://doi.org/10.1117/12.841169

TY - GEN

T1 - Regulatory assembly of the vacuolar proton pump VOV1-ATPase in yeast cells by FLIM-FRET

AU - Ernst, Stefan

AU - Batisse, Claire

AU - Zarrabi, Nawid

AU - Boettcher, Bettina

AU - Boersch, Michael

PY - 2010

Y1 - 2010

N2 - 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.

AB - 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.

U2 - 10.1117/12.841169

DO - 10.1117/12.841169

M3 - Conference contribution

SN - 978-0-8194-7965-5

SP - -

BT - MULTIPHOTON MICROSCOPY IN THE BIOMEDICAL SCIENCES X

A2 - Periasamy, A

A2 - So, PTC

A2 - Konig, K

PB - SPIE

CY - BELLINGHAM

T2 - Conference on Multiphoton Microscopy in the Biomedical Sciences X

Y2 - 24 January 2010 through 26 January 2010

ER -

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

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