title: 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.
authors.affiliation: Department of Cell Physiology and Molecular Biophysics and Center for Membrane Protein Research, Texas Tech University Health Sciences Center, 3601 4th Street STOP 6551, Lubbock, TX 79430, USA.
authors.forename: Cholpon
authors.initials: C
authors.lastname: Tilegenova
authors.affiliation: Current address: Department of Biology, Texas State University, 601 University Drive, San Marcos, TX 78666, USA.
authors.forename: Spandana
authors.initials: S
authors.lastname: Vemulapally
authors.affiliation: Department of Cell Physiology and Molecular Biophysics and Center for Membrane Protein Research, Texas Tech University Health Sciences Center, 3601 4th Street STOP 6551, Lubbock, TX 79430, USA.
authors.forename: Doris M
authors.initials: DM
authors.lastname: Cortes
authors.affiliation: Department of Cell Physiology and Molecular Biophysics and Center for Membrane Protein Research, Texas Tech University Health Sciences Center, 3601 4th Street STOP 6551, Lubbock, TX 79430, USA. Electronic address: luis.cuello@ttuhsc.edu.
authors.forename: Luis G
authors.initials: LG
authors.lastname: Cuello
mesh_terms: D001426:Bacterial Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D015221:Potassium Channels / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D048170:Streptomyces lividans / Q000235:genetics* / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D015221:Potassium Channels; D011994:Recombinant Proteins
keywords: Biochemistry; Detergents; Ion channels; KcsA; Over-expression; Solubilization