Dhillon, MS, Cockcroft, CJ, Munsey, T et al. (7 more authors) (2014) A functional Kv1.2-hERG chimaeric channel expressed in Pichia pastoris. Scientific Reports, 4. 4201. ISSN 2045-2322
Abstract
Members of the six-transmembrane segment family of ion channels share a common structural design. However, there are sequence differences between the members that confer distinct biophysical properties on individual channels. Currently, we do not have 3D structures for all members of the family to help explain the molecular basis for the differences in their biophysical properties and pharmacology. This is due to low-level expression of many members in native or heterologous systems. One exception is rat Kv1.2 which has been overexpressed in Pichia pastoris and crystallised. Here, we tested chimaeras of rat Kv1.2 with the hERG channel for function in Xenopus oocytes and for overexpression in Pichia. Chimaera containing the S1-S6 transmembrane region of HERG showed functional and pharmacological properties similar to hERG and could be overexpressed and purified from Pichia. Our results demonstrate that rat Kv1.2 could serve as a surrogate to express difficult-to-overexpress members of the six-transmembrane segment channel family.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | (c) 2014, The Author(s). This is an open access article under the terms of the Creative Commons Attribution CC BY 3.0 License |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Medicine and Health (Leeds) > School of Medicine (Leeds) > Inst of Biomed & Clin Sciences (LIBACS) (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 22 Aug 2014 12:50 |
Last Modified: | 22 Aug 2014 12:50 |
Published Version: | http://dx.doi.org/10.1038/srep04201 |
Status: | Published |
Publisher: | Nature Publishing Group |
Identification Number: | 10.1038/srep04201 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:80031 |