Maue Robert A
Department of Physiology, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
J Cell Physiol. 2007 Dec;213(3):618-25. doi: 10.1002/jcp.21259.
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.
表位标签已越来越多地用于了解离子通道亚基的组装与相互作用、运输、亚细胞定位以及在活细胞中的功能。特别是，表位标签已被证明对于在体外和体内内源性细胞环境中分析密切相关、高度同源的通道亚基极为有用，因为在这些环境中可能会表达多种通道亚型。然而，随着所使用的表位标签种类不断增加，并且标记通道亚基的使用变得越来越复杂和广泛，突出与在离子通道研究中使用表位标签相关潜在问题的例子数量也有所增加。本文介绍了一些用于研究离子通道亚基的表位标签，包括HA、FLAG、myc、His6和绿色荧光蛋白（GFP）表位，以及它们已被证明具有优势的一些应用和研究途径。还讨论了与使用这些表位标签相关的潜在陷阱和注意事项，重点强调需要将仔细表征表位标记的通道亚基作为其构建的一部分。最后，考虑了未来研究和该方法发展的潜在途径。
