Membrane protein expression and analysis in yeast
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Published on: June 13, 2021
Katrien Claes1, Mouna Guerfal1, Nico Callewaert1
1Unit 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.
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.
