title: Use of Escherichia coli for the production and purification of membrane proteins.
abstract: Individual types of ion channels and other membrane proteins are typically expressed only at low levels in their native membranes, rendering their isolation by conventional purification techniques difficult. The heterologous over-expression of such proteins is therefore usually a prerequisite for their purification in amounts suitable for structural and for many functional investigations. The most straightforward expression host, suitable for prokaryote membrane proteins and some proteins from eukaryotes, is the bacterium Escherichia coli. Here we describe the use of this expression system for production of functionally active polytopic membrane proteins and methods for their purification by affinity chromatography in amounts up to tens of milligrams.
authors.affiliation: Astbury Centre for Structural Molecular Biology, School of Biomedical Sciences, University of Leeds, Leeds, UK.
authors.forename: Vincent G L
authors.initials: VG
authors.lastname: Postis
authors.forename: Andrea E
authors.initials: AE
authors.lastname: Rawlings
authors.forename: Amelia
authors.initials: A
authors.lastname: Lesiuk
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Baldwin
mesh_terms: D015153:Blotting, Western; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D007473:Ion Channels / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D026901:Membrane Transport Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid
chemical_list: D007473:Ion Channels; D008565:Membrane Proteins; D026901:Membrane Transport Proteins