Fluorescence resonance energy transfer analysis of subunit assembly of the ASIC channel

Ying Gao, Shuang Shuang Liu, Shuang Qiu, Wei Cheng, Jie Zheng, Jian Hong Luo

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Acid-sensing ion channels (ASICs) are believed to be homo- or heteromeric complexes, which have been verified by classical methods such as co-immunoprecipitation or electrophysiological assays. However, the exact subunit combinations of ASICs in living cells have not been established yet. Here, we apply assays based on fluorescence resonance energy transfer (FRET) between GFP color mutants CFP and YFP to investigate ASIC assembly directly in living cells. Homomerization as well as heteromerization of different combinations of ASIC subunits were found. In addition, our results suggest the formation of heteromeric 1a/2a channels of stoichiometry consisting of at least two 1a subunits and two 2a subunits. Similar stoichiometry was observed from heteromeric 1a/2b and 2a/2b channels. Our results imply that these heteromeric ASIC channels contain at least four subunits.

Original languageEnglish (US)
Pages (from-to)143-150
Number of pages8
JournalBiochemical and Biophysical Research Communications
Volume359
Issue number1
DOIs
StatePublished - Jul 20 2007

Keywords

  • ASIC
  • Assembly
  • CHO
  • FRET
  • Heteromer
  • Homomer
  • Stoichiometry

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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