Atomic force microscopy imaging reveals the formation of ASIC/ENaC cross-clade ion channels
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Peer-reviewed
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Abstract
ASIC and ENaC are co-expressed in various cell types, and there is evidence for a close association between them. Here, we used atomic force microscopy (AFM) to determine whether ASIC1a and ENaC subunits are able to form cross-clade hybrid ion channels. ASIC1a and ENaC could be co-isolated from detergent extracts of tsA 201 cells co-expressing the two subunits. Isolated proteins were incubated with antibodies against ENaC and Fab fragments against ASIC1a. AFM imaging revealed proteins that were decorated by both an antibody and a Fab fragment with an angle of ∼120° between them, indicating the formation of ASIC1a/ENaC heterotrimers.
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Keywords
Acid-sensing ion channel (ASIC), Atomic force microscopy, Channel assembly, Epithelial Na(+) channel (ENaC), Acid Sensing Ion Channels, Animals, Antibodies, CHO Cells, Cell Line, Transformed, Cricetulus, Epithelial Sodium Channels, Epitopes, Gene Expression, HEK293 Cells, Humans, Hydrogen-Ion Concentration, Microscopy, Atomic Force, Patch-Clamp Techniques, Protein Multimerization, Proto-Oncogene Proteins c-myc, Recombinant Fusion Proteins
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Biochemical and Biophysical Research Communications
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0006-291X
1090-2104
1090-2104
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464
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Elsevier
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J.M.E. and P.J. are supported by Kidney Research U.K. A.P.S. was a member of the University of Cambridge MB/PhD Programme, and was supported by the Jean Shanks Foundation and the James Baird Fund. A research stay of S.H. at Cambridge was supported by a PhD fellowship from the Bayerische Forschungsstiftung.