title: A cost-effective protocol for the over-expression and purification of fully-functional and more stable Erwinia chrysanthemi ligand-gated ion channel.
abstract: The Erwinia chrysanthemi ligand-gated ion channel, ELIC, is considered an excellent structural and functional surrogate for the whole pentameric ligand-gated ion channel family. Despite its simplicity, ELIC is structurally capable of undergoing ligand-dependent activation and a concomitant desensitization process. To determine at the molecular level the structural changes underlying ELIC's function, it is desirable to produce large quantities of protein. This protein should be properly folded, fully-functional and amenable to structural determinations. In the current paper, we report a completely new protocol for the expression and purification of milligram quantities of fully-functional, more stable and crystallizable ELIC. The use of an autoinduction media and inexpensive detergents during ELIC extraction, in addition to the high-quality and large quantity of the purified channel, are the highlights of this improved biochemical protocol.
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: Benjamin W
authors.initials: BW
authors.lastname: Elberson
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: Ty E
authors.initials: TE
authors.lastname: Whisenant
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: D Marien
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 / Q000737:chemistry* / Q000302:isolation & purification*; D016974:Dickeya chrysanthemi / Q000737:chemistry*; D058446:Ligand-Gated Ion Channels / Q000737:chemistry* / Q000302:isolation & purification*
chemical_list: D001426:Bacterial Proteins; D058446:Ligand-Gated Ion Channels