Membrane protein expression and analysis in yeast.

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.