Purification of the functional plant membrane channel KAT1

Takao Hibi, Shiho Aoki, Keisuke Oda, Shintaro Munemasa, Shunsuke Ozaki, Osamu Shirai, Yoshiyuki Murata, Nobuyuki Uozumi

Research output: Contribution to journalArticle

5 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)465-469
Number of pages5
JournalBiochemical and Biophysical Research Communications
Volume374
Issue number3
DOIs
Publication statusPublished - Sep 26 2008

Fingerprint

Ion Channels
Detergents
Purification
Chromatography
Arabidopsis
Gel Chromatography
Membrane Proteins
Fusion reactions
Gels
Availability
Proteins

Keywords

  • Affinity chromatography
  • Detergent screening
  • Functional purification
  • Green fluorescent protein
  • Membrane protein
  • Potassium channel

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Purification of the functional plant membrane channel KAT1. / Hibi, Takao; Aoki, Shiho; Oda, Keisuke; Munemasa, Shintaro; Ozaki, Shunsuke; Shirai, Osamu; Murata, Yoshiyuki; Uozumi, Nobuyuki.

In: Biochemical and Biophysical Research Communications, Vol. 374, No. 3, 26.09.2008, p. 465-469.

Research output: Contribution to journalArticle

Hibi, Takao ; Aoki, Shiho ; Oda, Keisuke ; Munemasa, Shintaro ; Ozaki, Shunsuke ; Shirai, Osamu ; Murata, Yoshiyuki ; Uozumi, Nobuyuki. / Purification of the functional plant membrane channel KAT1. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 374, No. 3. pp. 465-469.
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