Calmodulin interaction with heag1 visualized by fret microscopy

J. Tiago Goncalves, Walter Stühmer

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Background: Ca2+-mediated regulation of ion channels provides a link between intracellular signaling pathways and membrane electrical activity. Intracellular Ca2+ inhibits the voltage-gated potassium channel EAG1 through the direct binding of calmodulin (CaM). Three CaM binding sites (BD-C1: 674-683, BD-C2: 711-721, BD-N: 151-165) have been identified in a peptide screen and were proposed to mediate binding. The participation of the three sites in CaM binding to the native channel, however, remains unclear. Methodology/Principal Findings: Here we studied the binding of Ca2+/CaM to the EAG channel by visualizing the interaction between YFP-labeled CaM and Cerulean-labeled hEAG1 in mammalian cells by FRET. The results of our cellular approach substantiate that two CaM binding sites are predominantly involved; the high-affinity 1-8-14 based CaM binding domain in the N-terminus and the second C-terminal binding domain BD-C2. Mutations at these sites completely abolished CaM binding to hEAG1. Conclusions/Significance: We demonstrated that the BD-N and BD-C2 binding domains are sufficient for CaM binding to the native channel, and, therefore, that BD-C1 is unable to bind CaM independently.

Original languageEnglish (US)
Article numbere10873
JournalPLoS One
Volume5
Issue number5
DOIs
StatePublished - 2010
Externally publishedYes

Fingerprint

calmodulin
Calmodulin
Microscopy
microscopy
Microscopic examination
calcium
binding sites
Binding Sites
Voltage-Gated Potassium Channels
potassium channels
ion channels
Ion Channels
Cells
peptides
Membranes
mutation
Peptides
Mutation

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Calmodulin interaction with heag1 visualized by fret microscopy. / Goncalves, J. Tiago; Stühmer, Walter.

In: PLoS One, Vol. 5, No. 5, e10873, 2010.

Research output: Contribution to journalArticle

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