title: Membrane protein expression and analysis in yeast.
abstract: This protocol describes the expression and analysis of membrane proteins produced in yeast, as illustrated with Yarrowia lipolytica and Pichia pastoris. Step by step, we explain how to generate a yeast strain expressing the membrane protein of interest, how to prepare a membrane protein sample from yeast, and how to analyze the expression levels using SDS-PAGE and Western blotting. In the final section, we describe how to perform a radioligand binding assay, which quantifies the amount of the protein folded in a ligand-binding competent state.
authors: Unit of Medical Biotechnology, Department of Medical Protein Research, VIB-UGhent, Ghent, Belgium; Inflammation Research Center, VIB-UGhent, Ghent, Belgium; Department of Biochemistry and Microbiology, Laboratory for Protein Biochemistry and Biomolecular Engineering, Ghent University, Ghent, Belgium.
authors: Katrien
authors: K
authors: Claes
authors: Unit of Medical Biotechnology, Department of Medical Protein Research, VIB-UGhent, Ghent, Belgium; Inflammation Research Center, VIB-UGhent, Ghent, Belgium; Department of Biochemistry and Microbiology, Laboratory for Protein Biochemistry and Biomolecular Engineering, Ghent University, Ghent, Belgium.
authors: Mouna
authors: M
authors: Guerfal
authors: Unit of Medical Biotechnology, Department of Medical Protein Research, VIB-UGhent, Ghent, Belgium; Inflammation Research Center, VIB-UGhent, Ghent, Belgium; Department of Biochemistry and Microbiology, Laboratory for Protein Biochemistry and Biomolecular Engineering, Ghent University, Ghent, Belgium. Electronic address: nico.callewaert@vib-ugent.be.
authors: Nico
authors: N
authors: Callewaert
mesh_terms: D015870:Gene Expression; D008565:Membrane Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D010843:Pichia / Q000235:genetics*; D014170:Transformation, Genetic; D025062:Yarrowia / Q000235:genetics*
chemical_list: D008565:Membrane Proteins
keywords: Membrane protein overexpression; Membrane protein preparation; Membrane proteins; Pichia pastoris; Radioligand binding assay; Yarrowia lipolytica; Yeast transformation