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Title: Atomic force microscopy imaging reveals the formation of ASIC/ENaC cross-clade ion channels

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. - Highlights: • There is evidence for a close association between ASIC and ENaC. • We used AFM to test whether ASIC1a and ENaC subunits form cross-clade ion channels. • Isolated proteins were incubated with subunit-specific antibodies and Fab fragments. • Some proteins were doubly decorated at ∼120° by an antibody and a Fab fragment. • Our results indicate the formation of ASIC1a/ENaC heterotrimers.

Authors:
; ;  [1]; ;  [2];  [1]
  1. Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD (United Kingdom)
  2. Institut für Zelluläre und Molekulare Physiologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Waldstrasse 6, 91054 Erlangen (Germany)
Publication Date:
OSTI Identifier:
22462174
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 464; Journal Issue: 1; Other Information: Copyright (c) 2015 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ANTIBODIES; ATOMIC FORCE MICROSCOPY; BIOMEDICAL RADIOGRAPHY; DETERGENTS; FLUORESCEIN; HAMSTERS; HYBRIDIZATION; ISOTHIOCYANATES; KIDNEYS; MUSCLES; OVARIES; PROTEINS

Citation Formats

Jeggle, Pia, Smith, Ewan St. J., Stewart, Andrew P., Haerteis, Silke, Korbmacher, Christoph, and Edwardson, J. Michael, E-mail: jme1000@cam.ac.uk. Atomic force microscopy imaging reveals the formation of ASIC/ENaC cross-clade ion channels. United States: N. p., 2015. Web. doi:10.1016/J.BBRC.2015.05.091.
Jeggle, Pia, Smith, Ewan St. J., Stewart, Andrew P., Haerteis, Silke, Korbmacher, Christoph, & Edwardson, J. Michael, E-mail: jme1000@cam.ac.uk. Atomic force microscopy imaging reveals the formation of ASIC/ENaC cross-clade ion channels. United States. doi:10.1016/J.BBRC.2015.05.091.
Jeggle, Pia, Smith, Ewan St. J., Stewart, Andrew P., Haerteis, Silke, Korbmacher, Christoph, and Edwardson, J. Michael, E-mail: jme1000@cam.ac.uk. Fri . "Atomic force microscopy imaging reveals the formation of ASIC/ENaC cross-clade ion channels". United States. doi:10.1016/J.BBRC.2015.05.091.
@article{osti_22462174,
title = {Atomic force microscopy imaging reveals the formation of ASIC/ENaC cross-clade ion channels},
author = {Jeggle, Pia and Smith, Ewan St. J. and Stewart, Andrew P. and Haerteis, Silke and Korbmacher, Christoph and Edwardson, J. Michael, E-mail: jme1000@cam.ac.uk},
abstractNote = {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. - Highlights: • There is evidence for a close association between ASIC and ENaC. • We used AFM to test whether ASIC1a and ENaC subunits form cross-clade ion channels. • Isolated proteins were incubated with subunit-specific antibodies and Fab fragments. • Some proteins were doubly decorated at ∼120° by an antibody and a Fab fragment. • Our results indicate the formation of ASIC1a/ENaC heterotrimers.},
doi = {10.1016/J.BBRC.2015.05.091},
journal = {Biochemical and Biophysical Research Communications},
number = 1,
volume = 464,
place = {United States},
year = {Fri Aug 14 00:00:00 EDT 2015},
month = {Fri Aug 14 00:00:00 EDT 2015}
}
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