title: Expression of Eukaryotic Membrane Proteins in Pichia pastoris.
abstract: A key point when it comes to heterologous expression of eukaryotic membrane proteins (EMPs) is the choice of the best-suited expression platform. The yeast Pichia pastoris has proven to be a very versatile system showing promising results in a growing number of cases. Indeed, its particular methylotrophic characteristics combined to the very simple handling of a eukaryotic microorganism that possesses the majority of mammalian-like machineries make it a very competitive expression system for various complex proteins, in amounts compatible with functional and structural studies. This chapter describes a set of robust methodologies routinely used for the successful expression of a variety of EMPs, going from yeast transformation with the recombinant plasmid to the analysis of the quality and quantity of the proteins produced.
authors.affiliation: Biotechnology and Cell Signalling, IMPReSs Protein Facility, UMR7242 ESBS CNRS, University of Strasbourg, 300 Blvd S. Brant, 67412, Illkirch, France.
authors.forename: Lucie
authors.initials: L
authors.lastname: Hartmann
authors.affiliation: Biotechnology and Cell Signalling, IMPReSs Protein Facility, UMR7242 ESBS CNRS, University of Strasbourg, 300 Blvd S. Brant, 67412, Illkirch, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Kugler
authors.affiliation: Biotechnology and Cell Signalling, IMPReSs Protein Facility, UMR7242 ESBS CNRS, University of Strasbourg, 300 Blvd S. Brant, 67412, Illkirch, France. renaud.wagner@unistra.fr.
authors.forename: Renaud
authors.initials: R
authors.lastname: Wagner
mesh_terms: D003001:Cloning, Molecular; D005057:Eukaryotic Cells / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism*; D010843:Pichia / Q000235:genetics / Q000254:growth & development*; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D008565:Membrane Proteins; D011994:Recombinant Proteins
keywords: Eukaryotic membrane protein; GPCR; Heterologous expression; Pichia pastoris; Yeast