Biophysical characterization of the fluorescent protein voltage probe VSFP2.3 based on the voltage-sensing domain of Ci-VSP

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Biophysical characterization of the fluorescent protein voltage probe VSFP2.3 based on the voltage-sensing domain of Ci-VSP. / Lundby, Alicia; Akemann, Walther; Knöpfel, Thomas.

In: European Biophysics Journal, Vol. 39, No. 12, 06.08.2010, p. 1625-1635.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Lundby, A, Akemann, W & Knöpfel, T 2010, 'Biophysical characterization of the fluorescent protein voltage probe VSFP2.3 based on the voltage-sensing domain of Ci-VSP', European Biophysics Journal, vol. 39, no. 12, pp. 1625-1635. https://doi.org/10.1007/s00249-010-0620-0

APA

Lundby, A., Akemann, W., & Knöpfel, T. (2010). Biophysical characterization of the fluorescent protein voltage probe VSFP2.3 based on the voltage-sensing domain of Ci-VSP. European Biophysics Journal, 39(12), 1625-1635. https://doi.org/10.1007/s00249-010-0620-0

Vancouver

Lundby A, Akemann W, Knöpfel T. Biophysical characterization of the fluorescent protein voltage probe VSFP2.3 based on the voltage-sensing domain of Ci-VSP. European Biophysics Journal. 2010 Aug 6;39(12):1625-1635. https://doi.org/10.1007/s00249-010-0620-0

Author

Lundby, Alicia ; Akemann, Walther ; Knöpfel, Thomas. / Biophysical characterization of the fluorescent protein voltage probe VSFP2.3 based on the voltage-sensing domain of Ci-VSP. In: European Biophysics Journal. 2010 ; Vol. 39, No. 12. pp. 1625-1635.

Bibtex

@article{ffca5961b79a4596b4fc0d201109c755,
title = "Biophysical characterization of the fluorescent protein voltage probe VSFP2.3 based on the voltage-sensing domain of Ci-VSP",
abstract = "A voltage sensitive phosphatase was discovered in the ascidian Ciona intestinalis. The phosphatase, Ci-VSP, contains a voltage-sensing domain homologous to those known from voltage-gated ion channels, but unlike ion channels, the voltage-sensing domain of Ci-VSP can reside in the cell membrane as a monomer. We fused the voltage-sensing domain of Ci-VSP to a pair of fluorescent reporter proteins to generate a genetically encodable voltage-sensing fluorescent probe, VSFP2.3. VSFP2.3 is a fluorescent voltage probe that reports changes in membrane potential as a FRET (fluorescence resonance energy transfer) signal. Here we report sensing current measurements from VSFP2.3, and show that VSFP2.3 carries 1.2 e sensing charges, which are displaced within 1.5 ms. The sensing currents become faster at higher temperatures, and the voltage dependence of the decay time constants is temperature dependent. Neutralization of an arginine in S4, previously suggested to be a sensing charge, and measuring associated sensing currents indicate that this charge is likely to reside at the membrane-aqueous interface rather than within the membrane electric field. The data presented give us insights into the voltage-sensing mechanism of Ci-VSP, which will allow us to further improve the sensitivity and kinetics of the family of VSFP proteins.",
keywords = "Animals, Arginine, Cell Membrane, Electric Conductivity, Electrophysiology, Fluorescence Resonance Energy Transfer, Ion Channel Gating, Ion Channels, Kinetics, Luminescent Proteins, Membrane Potentials, PC12 Cells, Phosphoric Monoester Hydrolases, Rats, Temperature",
author = "Alicia Lundby and Walther Akemann and Thomas Kn{\"o}pfel",
year = "2010",
month = "8",
day = "6",
doi = "10.1007/s00249-010-0620-0",
language = "English",
volume = "39",
pages = "1625--1635",
journal = "European Biophysics Journal",
issn = "0175-7571",
publisher = "Springer",
number = "12",

}

RIS

TY - JOUR

T1 - Biophysical characterization of the fluorescent protein voltage probe VSFP2.3 based on the voltage-sensing domain of Ci-VSP

AU - Lundby, Alicia

AU - Akemann, Walther

AU - Knöpfel, Thomas

PY - 2010/8/6

Y1 - 2010/8/6

N2 - A voltage sensitive phosphatase was discovered in the ascidian Ciona intestinalis. The phosphatase, Ci-VSP, contains a voltage-sensing domain homologous to those known from voltage-gated ion channels, but unlike ion channels, the voltage-sensing domain of Ci-VSP can reside in the cell membrane as a monomer. We fused the voltage-sensing domain of Ci-VSP to a pair of fluorescent reporter proteins to generate a genetically encodable voltage-sensing fluorescent probe, VSFP2.3. VSFP2.3 is a fluorescent voltage probe that reports changes in membrane potential as a FRET (fluorescence resonance energy transfer) signal. Here we report sensing current measurements from VSFP2.3, and show that VSFP2.3 carries 1.2 e sensing charges, which are displaced within 1.5 ms. The sensing currents become faster at higher temperatures, and the voltage dependence of the decay time constants is temperature dependent. Neutralization of an arginine in S4, previously suggested to be a sensing charge, and measuring associated sensing currents indicate that this charge is likely to reside at the membrane-aqueous interface rather than within the membrane electric field. The data presented give us insights into the voltage-sensing mechanism of Ci-VSP, which will allow us to further improve the sensitivity and kinetics of the family of VSFP proteins.

AB - A voltage sensitive phosphatase was discovered in the ascidian Ciona intestinalis. The phosphatase, Ci-VSP, contains a voltage-sensing domain homologous to those known from voltage-gated ion channels, but unlike ion channels, the voltage-sensing domain of Ci-VSP can reside in the cell membrane as a monomer. We fused the voltage-sensing domain of Ci-VSP to a pair of fluorescent reporter proteins to generate a genetically encodable voltage-sensing fluorescent probe, VSFP2.3. VSFP2.3 is a fluorescent voltage probe that reports changes in membrane potential as a FRET (fluorescence resonance energy transfer) signal. Here we report sensing current measurements from VSFP2.3, and show that VSFP2.3 carries 1.2 e sensing charges, which are displaced within 1.5 ms. The sensing currents become faster at higher temperatures, and the voltage dependence of the decay time constants is temperature dependent. Neutralization of an arginine in S4, previously suggested to be a sensing charge, and measuring associated sensing currents indicate that this charge is likely to reside at the membrane-aqueous interface rather than within the membrane electric field. The data presented give us insights into the voltage-sensing mechanism of Ci-VSP, which will allow us to further improve the sensitivity and kinetics of the family of VSFP proteins.

KW - Animals

KW - Arginine

KW - Cell Membrane

KW - Electric Conductivity

KW - Electrophysiology

KW - Fluorescence Resonance Energy Transfer

KW - Ion Channel Gating

KW - Ion Channels

KW - Kinetics

KW - Luminescent Proteins

KW - Membrane Potentials

KW - PC12 Cells

KW - Phosphoric Monoester Hydrolases

KW - Rats

KW - Temperature

U2 - 10.1007/s00249-010-0620-0

DO - 10.1007/s00249-010-0620-0

M3 - Journal article

C2 - 20686764

VL - 39

SP - 1625

EP - 1635

JO - European Biophysics Journal

JF - European Biophysics Journal

SN - 0175-7571

IS - 12

ER -

ID: 45558648