High-Sensitivity Fluorometry to Resolve Ion Channel Conformational Dynamics

Department of Drug Design and Pharmacology

High-Sensitivity Fluorometry to Resolve Ion Channel Conformational Dynamics

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

High-Sensitivity Fluorometry to Resolve Ion Channel Conformational Dynamics. / Wulf, Matthias; Pless, Stephan Alexander.

In: Cell Reports, Vol. 22, No. 6, 06.02.2018, p. 1615-1626.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Wulf, M & Pless, SA 2018, 'High-Sensitivity Fluorometry to Resolve Ion Channel Conformational Dynamics', Cell Reports, vol. 22, no. 6, pp. 1615-1626. https://doi.org/10.1016/j.celrep.2018.01.029

APA

Wulf, M., & Pless, S. A. (2018). High-Sensitivity Fluorometry to Resolve Ion Channel Conformational Dynamics. Cell Reports, 22(6), 1615-1626. https://doi.org/10.1016/j.celrep.2018.01.029

Vancouver

Wulf M, Pless SA. High-Sensitivity Fluorometry to Resolve Ion Channel Conformational Dynamics. Cell Reports. 2018 Feb 6;22(6):1615-1626. https://doi.org/10.1016/j.celrep.2018.01.029

Author

Wulf, Matthias ; Pless, Stephan Alexander. / High-Sensitivity Fluorometry to Resolve Ion Channel Conformational Dynamics. In: Cell Reports. 2018 ; Vol. 22, No. 6. pp. 1615-1626.

Bibtex

@article{73eac2d10594469880f9a9041de51389,

title = "High-Sensitivity Fluorometry to Resolve Ion Channel Conformational Dynamics",

abstract = "Fluorescent labels offer the capability to follow conformational dynamics of membrane proteins, but signal detection in such recordings is inherently difficult to achieve in a cell membrane and lacks sufficient time resolution to follow physiologically relevant transitions. Here, we develop high-sensitivity patch-clamp fluorometry (hsPCF), a fluorescence-based approach that results in up to 10-fold increased signals and affords 50-fold faster fluorescence recordings than previous methods. The increased time resolution is paired with a very high versatility in terms of the choice of fluorescent dye, cell type, and protein of interest. We highlight this versatility by providing insight into the conformational dynamics of both ligand- and voltage-gated ion channels using fluorescent labels introduced in extracellular or transmembrane positions while changing either the extra- or intracellular solutions. Together, hsPCF will thus enable the future study of membrane-embedded proteins with sufficient temporal resolution to resolve conformational dynamics.",

keywords = "Journal Article",

author = "Matthias Wulf and Pless, {Stephan Alexander}",

year = "2018",

month = feb,

day = "6",

doi = "10.1016/j.celrep.2018.01.029",

language = "English",

volume = "22",

pages = "1615--1626",

journal = "Cell Reports",

issn = "2211-1247",

publisher = "Cell Press",

number = "6",

}

RIS

TY - JOUR

T1 - High-Sensitivity Fluorometry to Resolve Ion Channel Conformational Dynamics

AU - Wulf, Matthias

AU - Pless, Stephan Alexander

PY - 2018/2/6

Y1 - 2018/2/6

N2 - Fluorescent labels offer the capability to follow conformational dynamics of membrane proteins, but signal detection in such recordings is inherently difficult to achieve in a cell membrane and lacks sufficient time resolution to follow physiologically relevant transitions. Here, we develop high-sensitivity patch-clamp fluorometry (hsPCF), a fluorescence-based approach that results in up to 10-fold increased signals and affords 50-fold faster fluorescence recordings than previous methods. The increased time resolution is paired with a very high versatility in terms of the choice of fluorescent dye, cell type, and protein of interest. We highlight this versatility by providing insight into the conformational dynamics of both ligand- and voltage-gated ion channels using fluorescent labels introduced in extracellular or transmembrane positions while changing either the extra- or intracellular solutions. Together, hsPCF will thus enable the future study of membrane-embedded proteins with sufficient temporal resolution to resolve conformational dynamics.

AB - Fluorescent labels offer the capability to follow conformational dynamics of membrane proteins, but signal detection in such recordings is inherently difficult to achieve in a cell membrane and lacks sufficient time resolution to follow physiologically relevant transitions. Here, we develop high-sensitivity patch-clamp fluorometry (hsPCF), a fluorescence-based approach that results in up to 10-fold increased signals and affords 50-fold faster fluorescence recordings than previous methods. The increased time resolution is paired with a very high versatility in terms of the choice of fluorescent dye, cell type, and protein of interest. We highlight this versatility by providing insight into the conformational dynamics of both ligand- and voltage-gated ion channels using fluorescent labels introduced in extracellular or transmembrane positions while changing either the extra- or intracellular solutions. Together, hsPCF will thus enable the future study of membrane-embedded proteins with sufficient temporal resolution to resolve conformational dynamics.

KW - Journal Article

U2 - 10.1016/j.celrep.2018.01.029

DO - 10.1016/j.celrep.2018.01.029

M3 - Journal article

C2 - 29425514

VL - 22

SP - 1615

EP - 1626

JO - Cell Reports

JF - Cell Reports

SN - 2211-1247

IS - 6

ER -

ID: 189757717