Gating rearrangements in cyclic nucleotide-gated channels revealed by patch-clamp fluorometry

Jie Zheng, William N. Zagotta

Research output: Contribution to journalArticle

72 Citations (Scopus)

Abstract

Site-specific fluorescence recordings have shown great promise in understanding conformational changes in signaling proteins. The reported applications on ion channels have been limited to extracellular sites in whole oocyte preparations. We are now able to directly monitor gating movements of the intracellular domains of cyclic nucleotide-gated channels using simultaneous site-specific fluorescence recording and patch-clamp current recording from inside-out patches. Fluorescence signals were reliably observed when fluorophore was covalently attached to a site between the cyclic nucleotide-binding domain and the pore. While iodide, an anionic quencher, has a higher quenching efficiency in the channel's closed state, thallium ion, a cationic quencher, has a higher quenching efficiency in the open state. The state and charge dependence of quenching suggests movements of charged or dipolar residues near the fluorophore during CNG channel activation.

Original languageEnglish (US)
Pages (from-to)369-374
Number of pages6
JournalNeuron
Volume28
Issue number2
StatePublished - 2000
Externally publishedYes

Fingerprint

Cyclic Nucleotide-Gated Cation Channels
Fluorometry
Fluorescence
Thallium
Cyclic Nucleotides
Iodides
Ion Channels
Oocytes
Ions
Proteins

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Gating rearrangements in cyclic nucleotide-gated channels revealed by patch-clamp fluorometry. / Zheng, Jie; Zagotta, William N.

In: Neuron, Vol. 28, No. 2, 2000, p. 369-374.

Research output: Contribution to journalArticle

@article{baf2e31891a04f44b46e64596ecbed99,
title = "Gating rearrangements in cyclic nucleotide-gated channels revealed by patch-clamp fluorometry",
abstract = "Site-specific fluorescence recordings have shown great promise in understanding conformational changes in signaling proteins. The reported applications on ion channels have been limited to extracellular sites in whole oocyte preparations. We are now able to directly monitor gating movements of the intracellular domains of cyclic nucleotide-gated channels using simultaneous site-specific fluorescence recording and patch-clamp current recording from inside-out patches. Fluorescence signals were reliably observed when fluorophore was covalently attached to a site between the cyclic nucleotide-binding domain and the pore. While iodide, an anionic quencher, has a higher quenching efficiency in the channel's closed state, thallium ion, a cationic quencher, has a higher quenching efficiency in the open state. The state and charge dependence of quenching suggests movements of charged or dipolar residues near the fluorophore during CNG channel activation.",
author = "Jie Zheng and Zagotta, {William N.}",
year = "2000",
language = "English (US)",
volume = "28",
pages = "369--374",
journal = "Neuron",
issn = "0896-6273",
publisher = "Cell Press",
number = "2",

}

TY - JOUR

T1 - Gating rearrangements in cyclic nucleotide-gated channels revealed by patch-clamp fluorometry

AU - Zheng, Jie

AU - Zagotta, William N.

PY - 2000

Y1 - 2000

N2 - Site-specific fluorescence recordings have shown great promise in understanding conformational changes in signaling proteins. The reported applications on ion channels have been limited to extracellular sites in whole oocyte preparations. We are now able to directly monitor gating movements of the intracellular domains of cyclic nucleotide-gated channels using simultaneous site-specific fluorescence recording and patch-clamp current recording from inside-out patches. Fluorescence signals were reliably observed when fluorophore was covalently attached to a site between the cyclic nucleotide-binding domain and the pore. While iodide, an anionic quencher, has a higher quenching efficiency in the channel's closed state, thallium ion, a cationic quencher, has a higher quenching efficiency in the open state. The state and charge dependence of quenching suggests movements of charged or dipolar residues near the fluorophore during CNG channel activation.

AB - Site-specific fluorescence recordings have shown great promise in understanding conformational changes in signaling proteins. The reported applications on ion channels have been limited to extracellular sites in whole oocyte preparations. We are now able to directly monitor gating movements of the intracellular domains of cyclic nucleotide-gated channels using simultaneous site-specific fluorescence recording and patch-clamp current recording from inside-out patches. Fluorescence signals were reliably observed when fluorophore was covalently attached to a site between the cyclic nucleotide-binding domain and the pore. While iodide, an anionic quencher, has a higher quenching efficiency in the channel's closed state, thallium ion, a cationic quencher, has a higher quenching efficiency in the open state. The state and charge dependence of quenching suggests movements of charged or dipolar residues near the fluorophore during CNG channel activation.

UR - http://www.scopus.com/inward/record.url?scp=0033635163&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033635163&partnerID=8YFLogxK

M3 - Article

C2 - 11144348

AN - SCOPUS:0033635163

VL - 28

SP - 369

EP - 374

JO - Neuron

JF - Neuron

SN - 0896-6273

IS - 2

ER -