| Literature DB >> 20064617 |
Nazzareno D'Avanzo1, Wayland W L Cheng, Xiaobing Xia, Liang Dong, Pavel Savitsky, Colin G Nichols, Declan A Doyle.
Abstract
The inward rectifier family of potassium (KCNJ) channels regulate vital cellular processes including cell volume, electrical excitability, and insulin secretion. Dysfunction of different isoforms have been linked to numerous diseases including Bartter's, Andersen-Tawil, Smith-Magenis Syndromes, Type II diabetes mellitus, and epilepsy, making them important targets for therapeutic intervention. Using a family-based approach, we succeeded in expressing 10 of 11 human KCNJ channels tested in Saccharomyces cerevisiae. GFP-fusion proteins showed that these channels traffic correctly to the plasma-membrane suggesting that the protein is functional. A 2-step purification process can be used to purify the KCNJ channels to >95% purity in a mono-dispersed form. After incorporation into liposomes, (86)Rb(+) flux assays confirm the functionality of the purified proteins as inward rectifier potassium channels. Copyright (c) 2010 Elsevier Inc. All rights reserved.Entities:
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Year: 2010 PMID： 20064617 PMCID： PMC3245711 DOI： 10.1016/j.pep.2010.01.010
Source DB: PubMed Journal: Protein Expr Purif ISSN： 1046-5928 Impact factor: 1.650