期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 111, 期 22, 页码 8281-8286出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1324060111
关键词
single-molecule imaging; subunit counting; BLINaC; ENaC
资金
- Deutsche Forschungsgemeinschaft [GR1771/3-5]
- Excellence Initiative of the German Federal Government [EXC 294]
- Excellence Initiative of the German State Government [EXC 294]
Acid-sensing ion channels (ASICs) are widely expressed protongated Na+ channels playing a role in tissue acidosis and pain. A trimeric composition of ASICs has been suggested by crystallization. Upon coexpression of ASIC1a and ASIC2a in Xenopus oocytes, we observed the formation of heteromers and their coexistence with homomers by electrophysiology, but could not determine whether heteromeric complexes have a fixed subunit stoichiometry or whether certain stoichiometries are preferred over others. We therefore imaged ASICs labeled with green and red fluorescent proteins on a single-molecule level, counted bleaching steps from GFP and colocalized them with red tandem tetrameric mCherry for many individual complexes. Combinatorial analysis suggests a model of random mixing of ASIC1a and ASIC2a subunits to yield both 2:1 and 1:2 ASIC1a: ASIC2a heteromers together with ASIC1a and ASIC2a homomers.
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