Membrane protein expression and analysis in yeast.

Autor:	Claes K; 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., Guerfal M; 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., Callewaert N; 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.
Jazyk:	angličtina
Zdroj:	Methods in enzymology [Methods Enzymol] 2015; Vol. 556, pp. 123-40. Date of Electronic Publication: 2015 Mar 20.
DOI:	10.1016/bs.mie.2014.12.010
Abstrakt:	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. (© 2015 Elsevier Inc. All rights reserved.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu