title: MemStar: a one-shot Escherichia coli-based approach for high-level bacterial membrane protein production.
abstract: Optimising membrane protein production yields in Escherichiacoli can be time- and resource-consuming. Here, we present a simple and effective Membrane protein Single shot amplification recipe: MemStar. This one-shot amplification recipe is based on the E. coli strain Lemo21(DE3), the PASM-5052 auto-induction medium and, contradictorily, an IPTG induction step. Using MemStar, production yields for most bacterial membrane proteins tested were improved to reach an average of 5 mg L(-1) per OD600 unit, which is significantly higher than yields obtained with other common production strategies. With MemStar, we have been able to obtain new structural information for several transporters, including the sodium/proton antiporter NapA.
authors.affiliation: Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, UK.
authors.forename: Chiara
authors.initials: C
authors.lastname: Lee
authors.affiliation: Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, UK.
authors.forename: Hae Joo
authors.initials: HJ
authors.lastname: Kang
authors.affiliation: Centre for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
authors.forename: Anna
authors.initials: A
authors.lastname: Hjelm
authors.affiliation: Centre for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
authors.forename: Abdul Aziz
authors.initials: AA
authors.lastname: Qureshi
authors.affiliation: Centre for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Nji
authors.affiliation: Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, UK; Membrane Protein Laboratory, Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE, UK; Rutherford Appleton Laboratory, Research Complex at Harwell, Didcot, Oxfordshire OX11 0DE, UK.
authors.forename: Hassanul
authors.initials: H
authors.lastname: Choudhury
authors.affiliation: Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, UK; Membrane Protein Laboratory, Diamond Light Source, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0DE, UK; Rutherford Appleton Laboratory, Research Complex at Harwell, Didcot, Oxfordshire OX11 0DE, UK.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Beis
authors.affiliation: Centre for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden.
authors.forename: Jan-Willem
authors.initials: JW
authors.lastname: de Gier
authors.affiliation: Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, UK; Centre for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, SE-106 91 Stockholm, Sweden. Electronic address: ddrew@dbb.su.se.
authors.forename: David
authors.initials: D
authors.lastname: Drew
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002458:Cell Fractionation / Q000379:methods; D005591:Chemical Fractionation / Q000379:methods; D004926:Escherichia coli / Q000737:chemistry* / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007218:Industrial Microbiology / Q000379:methods*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins
keywords: Escherichia coli; High-throughput; Membrane protein production; X-ray crystallography