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An improved method for the cost-effective expression and purification of large quantities of KcsA.

ABSTRACT: KcsA, the bacterial K(+) channel from Streptomyces lividans, is the prototypical model system to study the functional and structural correlations of the pore domain of eukaryotic voltage-gated K(+) channels (Kv channels). It contains all the molecular elements responsible for ion conduction, activation, deactivation and inactivation gating [1]. KcsA's structural simplicity makes it highly amenable for structural studies. Therefore, it is methodological advantageous to produce large amounts of functional and properly folded KcsA in a cost-effective manner. In the present study, we show an optimized protocol for the over-expression and purification of large amounts of high-quality, fully functional and crystallizable KcsA using inexpensive detergents, which significantly lowered the cost of the purification process.

SUBMITTER: Tilegenova C

PROVIDER: S-EPMC5662131 | biostudies-literature | 2016 Nov

REPOSITORIES: biostudies-literature

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An improved method for the cost-effective expression and purification of large quantities of KcsA.

Tilegenova Cholpon C Vemulapally Spandana S Cortes Doris M DM Cuello Luis G LG

Protein expression and purification 20160705

KcsA, the bacterial K(+) channel from Streptomyces lividans, is the prototypical model system to study the functional and structural correlations of the pore domain of eukaryotic voltage-gated K(+) channels (Kv channels). It contains all the molecular elements responsible for ion conduction, activation, deactivation and inactivation gating [1]. KcsA's structural simplicity makes it highly amenable for structural studies. Therefore, it is methodological advantageous to produce large amounts of fu ...[more]

PMID: 27393071

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