title: Expression and purification of the recombinant membrane protein YidC: a case study for increased stability and solubility.
abstract: YidC is an inner membrane protein from Escherichia coli and is an essential component in insertion, translocation and assembly of membrane proteins in the membranes. Previous purification attempts resulted in heavy aggregates and precipitated protein at later stages of purification. Here we present a rapid and straightforward stability screening strategy based on gel filtration chromatography, which requires as little as 10 microg of protein and takes less than 15 min to perform. With this technique, we could rapidly screen several buffers in order to identify an optimum condition that stabilizes purified YidC. After optimization we could obtain several milligrams of purified YidC that could be easily prepared at high concentrations and that was stable for weeks at +4 degrees C. The isolated protein is thus well suited for structural studies.
authors.affiliation: Division of Biophysics, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Daniel
authors.initials: D
authors.lastname: Martinez Molina
authors.forename: Anna-Karin
authors.initials: AK
authors.lastname: Lundbäck
authors.forename: Damian
authors.initials: D
authors.lastname: Niegowski
authors.forename: Said
authors.initials: S
authors.lastname: Eshaghi
mesh_terms: D002021:Buffers; D015180:Electrophoresis, Gel, Two-Dimensional; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D015870:Gene Expression; D026901:Membrane Transport Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility
chemical_list: D002021:Buffers; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D011994:Recombinant Proteins; C412443:YIDC protein, E coli