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

Jie Zheng, William N. Zagotta

Research output: Contribution to journalArticle

76 Scopus citations

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

ASJC Scopus subject areas

  • Neuroscience(all)

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