title: The synthesis of recombinant membrane proteins in yeast for structural studies.
abstract: Historically, recombinant membrane protein production has been a major challenge meaning that many fewer membrane protein structures have been published than those of soluble proteins. However, there has been a recent, almost exponential increase in the number of membrane protein structures being deposited in the Protein Data Bank. This suggests that empirical methods are now available that can ensure the required protein supply for these difficult targets. This review focuses on methods that are available for protein production in yeast, which is an important source of recombinant eukaryotic membrane proteins. We provide an overview of approaches to optimize the expression plasmid, host cell and culture conditions, as well as the extraction and purification of functional protein for crystallization trials in preparation for structural studies.
authors.affiliation: Department of Pharmacology, University of Cambridge, Tennis Court Road, Cambridge CB2 1PD, UK; School of Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK.
authors.forename: Sarah J
authors.initials: SJ
authors.lastname: Routledge
authors.affiliation: School of Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK.
authors.forename: Lina
authors.initials: L
authors.lastname: Mikaliunaite
authors.affiliation: School of Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK.
authors.forename: Anjana
authors.initials: A
authors.lastname: Patel
authors.affiliation: School of Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK.
authors.forename: Michelle
authors.initials: M
authors.lastname: Clare
authors.affiliation: School of Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK.
authors.forename: Stephanie P
authors.initials: SP
authors.lastname: Cartwright
authors.affiliation: School of Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK.
authors.forename: Zharain
authors.initials: Z
authors.lastname: Bawa
authors.affiliation: Smallpeice Enterprises Ltd, 27 Newbold Terrace East, Leamington Spa, Warwickshire CV32 4ES, UK.
authors.forename: Martin D B
authors.initials: MD
authors.lastname: Wilks
authors.affiliation: School of Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK.
authors.forename: Floren
authors.initials: F
authors.lastname: Low
authors.affiliation: School of Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK.
authors.forename: David
authors.initials: D
authors.lastname: Hardy
authors.affiliation: School of Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK.
authors.forename: Alice J
authors.initials: AJ
authors.lastname: Rothnie
authors.affiliation: School of Life & Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET, UK. Electronic address: r.m.bill@aston.ac.uk.
authors.forename: Roslyn M
authors.initials: RM
authors.lastname: Bill
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular / Q000379:methods*; D016208:Databases, Factual; D015870:Gene Expression; D005822:Genetic Vectors / Q000737:chemistry* / Q000378:metabolism; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids / Q000737:chemistry* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D012995:Solubility
chemical_list: D008565:Membrane Proteins; D011994:Recombinant Proteins
keywords: Cholesterol; Expression plasmid vector; Membrane proteins; Promoter; Recombinant; SMALP; Yeast