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.
authors.affiliation: Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, United Kingdom.
authors.forename: Pia
authors.initials: P
authors.lastname: Jeggle
authors.affiliation: Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, United Kingdom.
authors.forename: Ewan St J
authors.initials: ES
authors.lastname: Smith
authors.affiliation: Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, United Kingdom.
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Stewart
authors.affiliation: Institut für Zelluläre und Molekulare Physiologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Waldstrasse 6, 91054 Erlangen, Germany.
authors.forename: Silke
authors.initials: S
authors.lastname: Haerteis
authors.affiliation: Institut für Zelluläre und Molekulare Physiologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Waldstrasse 6, 91054 Erlangen, Germany.
authors.forename: Christoph
authors.initials: C
authors.lastname: Korbmacher
authors.affiliation: Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, United Kingdom. Electronic address: jme1000@cam.ac.uk.
authors.forename: J Michael
authors.initials: JM
authors.lastname: Edwardson
mesh_terms: D062565:Acid Sensing Ion Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000906:Antibodies / Q000737:chemistry; D016466:CHO Cells; D002461:Cell Line, Transformed; D003412:Cricetulus; D053503:Epithelial Sodium Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000939:Epitopes / Q000737:chemistry* / Q000378:metabolism; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018625:Microscopy, Atomic Force; D018408:Patch-Clamp Techniques; D055503:Protein Multimerization; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: C568266:ASIC1 protein, human; D062565:Acid Sensing Ion Channels; D000906:Antibodies; D053503:Epithelial Sodium Channels; D000939:Epitopes; C489427:MYC protein, human; D016271:Proto-Oncogene Proteins c-myc; D011993:Recombinant Fusion Proteins; C507045:SCNN1A protein, human; C507048:SCNN1B protein, human; C507052:SCNN1G protein, human
keywords: Acid-sensing ion channel (ASIC); Atomic force microscopy; Channel assembly; Epithelial Na(+) channel (ENaC)