Functional reconstitution of a prokaryotic K+ channel
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作者:
Heginbotham, L
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Brandeis Univ, Howard Hughes Med Inst, Dept Biochem, Waltham, MA 02254 USABrandeis Univ, Howard Hughes Med Inst, Dept Biochem, Waltham, MA 02254 USA
Heginbotham, L
[1
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Kolmakova-Partensky, L
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Brandeis Univ, Howard Hughes Med Inst, Dept Biochem, Waltham, MA 02254 USABrandeis Univ, Howard Hughes Med Inst, Dept Biochem, Waltham, MA 02254 USA
Kolmakova-Partensky, L
[1
]
Miller, C
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Brandeis Univ, Howard Hughes Med Inst, Dept Biochem, Waltham, MA 02254 USABrandeis Univ, Howard Hughes Med Inst, Dept Biochem, Waltham, MA 02254 USA
Miller, C
[1
]
机构:
[1] Brandeis Univ, Howard Hughes Med Inst, Dept Biochem, Waltham, MA 02254 USA
SliK, a K+ channel encoded by the Streptomyces KcsA gene, was expressed, purified, and reconstituted in liposomes. A concentrative Rb-86(+) flux assay was used to assess the ion transport properties of SliK. SliK-mediated ionic flux shows strong selectivity for K+ over Na+ and is inhibited by micromolar concentrations of Ba2+, mirroring the basic permeation characteristic of eukaryotic K+ channels studied by electrophysiological methods. Rb-86(+) uptake kinetics and equilibrium measurements also demonstrate that the purified protein is fully active.