Understanding ion channel biology using epitope tags: progress, pitfalls, and promise.
Epitope tags have been increasingly used to understand ion channel subunit assembly and interaction, trafficking, subcellular localization, and function in living cells. In particular, epitope tags have proven extremely useful for analyses of closely related, highly homologous channel subunits in endogenous cell contexts in vitro and in vivo, where multiple channel isoforms may be expressed. However, as the variety of epitope tags that have been used has expanded, and the use of tagged channel subunits has become increasingly sophisticated and widespread, there has also been an increase in the number of examples highlighting the potential problems associated with the use of epitope tags for ion channel studies. Described here are some of the epitope tags that have been used to study ion channel subunits, including the HA, FLAG, myc, His6, and green fluorescent protein (GFP) epitopes, as well as some of the applications and avenues of research in which they have proven advantageous. Potential pitfalls and caveats associated with the use of these epitope tags are also discussed, with an emphasis on the need to include careful characterization of epitope-tagged channel subunits as part of their construction. Finally, potential avenues for future investigation and the development of this approach are considered.
Maue
Robert A
RA
Department of Physiology, Dartmouth Medical School, Hanover, New Hampshire 03755, USA. Robert.Maue@Dartmouth.edu
D000818:Animals; D018604:Epitope Mapping; D000939:Epitopes / Q000235:genetics* / Q000378:metabolism*; D005544:Forecasting; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006388:Hemagglutinins / Q000378:metabolism; D006639:Histidine / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007473:Ion Channels / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D009842:Oligopeptides; D010455:Peptides / Q000378:metabolism; D016271:Proto-Oncogene Proteins c-myc / Q000378:metabolism
D000939:Epitopes; D006388:Hemagglutinins; D007473:Ion Channels; D009842:Oligopeptides; D010455:Peptides; D016271:Proto-Oncogene Proteins c-myc; D049452:Green Fluorescent Proteins; D006639:Histidine; C085706:FLAG peptide