Disruption of an intersubunit interaction underlies Ca 2+calmodulin modulation of cyclic nucleotide-gated channels

Jie Zheng, Michael D. Varnum, William N. Zagotta

Research output: Contribution to journalArticle

52 Scopus citations

Abstract

Cyclic nucleotide-gated channels are key molecular elements for olfactory transduction. Olfactory adaptation caused by repeated exposure to an odorant has been proposed to be mediated by the binding of Ca2+-calmodulin to the NH2-terminal domain of the channel, breaking its interaction with the COOH-terminal domain and downregulating the channel. We used a fluorescence resonance energy transfer (FRET) approach to study the structural aspects of this domain-domain interaction under physiological conditions in real time. Fluorescent proteins enhanced cyan fluorescent protein and enhanced yellow fluorescent protein were genetically attached at sites adjacent to the NH2- and COOH-terminal interacting domains, respectively, allowing direct observation of molecular rearrangements in intact channels. FRET signals caused by the specific interdomain interaction were observed in both intact cells and excised patches. Comparison of the effective FRET efficiencies demonstrated that the interaction occurs specifically between subunits but not within the same subunit. Binding of Ca2+-calmodulin caused a reversible decrease in FRET with the same time course as channel downregulation. These results suggest that a separation or reorientation of the interacting domains between subunits by Ca2+-calmodulin leads to channel downregulation. The quaternary arrangement presents a structural framework for understanding the molecular mechanism of olfactory adaptation.

Original languageEnglish (US)
Pages (from-to)8167-8175
Number of pages9
JournalJournal of Neuroscience
Volume23
Issue number22
StatePublished - Sep 3 2003
Externally publishedYes

Keywords

  • Ca-calmodulin
  • Fluorescence
  • FRET
  • GFP mutants
  • Ion channel
  • Olfactory adaptation
  • Signal transduction

ASJC Scopus subject areas

  • Neuroscience(all)

Fingerprint Dive into the research topics of 'Disruption of an intersubunit interaction underlies Ca <sup>2+</sup>calmodulin modulation of cyclic nucleotide-gated channels'. Together they form a unique fingerprint.

  • Cite this