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Purification of the functional plant membrane channel KAT1 |
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| Authors: | Hibi Takao Aoki Shiho Oda Keisuke Munemasa Shintaro Ozaki Shunsuke Shirai Osamu Murata Yoshiyuki Uozumi Nobuyuki |
| Institution: | a Department of Bioscience, Fukui Prefectural University, 4-1-1 MatsuokaKenjyojima, Eiheiji-cho, Fukui 910-1195, Japan b Department of Bioresources Chemistry, Okayama University, Tsushima-Naka, Okayama 700-8530, Japan c Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake, Sakyo-ku, Kyoto 606-8502, Japan d Department of Biomolecular Engineering, Graduate School of Engineering, Tohoku University, Aobayama 6-6-07, Sendai 980-8579, Japan |
| Abstract: | The inward-rectifying K+ channel KAT1 is expressed mainly in Arabidopsis thaliana guard cells. The purification of functional KAT1 has never been reported. We investigated the extraction of the plant K+ channel KAT1 with different detergents, as an example for how to select detergents for purifying a eukaryotic membrane protein. A KAT1-GFP fusion protein was used to screen a library of 46 detergents for the effective solubilization of intact KAT1. Then, a “test set” of three detergents was picked for further analysis, based on their biochemical characteristics and availability. The combination use of the selected detergents enabled the effective purification of functional KAT1 with affinity and gel-filtration chromatography. |
| Keywords: | DDM d-matopyranoside" target="_blank">n-dodecyl-β-d-matopyranoside NBT nitro-blue tetrazolium chloride BCIP 5-bromo-4-chloro-3&prime -indolylphosphatase p-toluidine salt |
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