Production of membrane proteins in yeast

Abstract

Background Yeast is an important and versatile organism for studying membrane proteins. It is easy to cultivate and can perform higher eukaryote-like post-translational modifications. S. cerevisiae has a fully-sequenced genome and there are several collections of deletion strains available, whilst P. pastoris can produce very high cell densities (230 g/l). Results We have used both S. cerevisiae and P. pastoris to over-produce the following His6 and His10 carboxyl terminal fused membrane proteins. CD81 – 26 kDa tetraspanin protein (TAPA-1) that may play an important role in the regulation of lymphoma cell growth and may also act as the viral receptor for Hepatitis C-Virus. CD82 – 30 kDa tetraspanin protein that associates with CD4 or CD8 cells and delivers co-stimulatory signals for the TCR/CD3 pathway. MC4R – 37 kDa seven transmembrane G-protein coupled receptor, present on neurons in the hypothalamus region of the brain and predicted to have a role in the feast or fast signalling pathway. Adt2p – 34 kDa six transmembrane protein that catalyses the exchange of ADP and ATP across the yeast mitochondrial inner membrane. Conclusion We show that yeasts are flexible production organisms for a range of different membrane proteins. The yields are such that future structure-activity relationship studies can be initiated via reconstitution, crystallization for X-ray diffraction or NMR experiments.

Publication DOI: https://doi.org/10.1186/1475-2859-5-S1-P38
Divisions: Life & Health Sciences > Biosciences
Life & Health Sciences
Life & Health Sciences > Cellular and Molecular Biomedicine
Additional Information: © Darby et al; licensee BioMed Central Ltd. 2006
Uncontrolled Keywords: yeast,membrane proteins,eukaryote-like post-translational modifications,S. cerevisiae,fully-sequenced genom,P. pastoris,cell densities,lymphoma cell growth,viral receptor,Hepatitis C-Virus,CD82 – 30 kDa tetraspanin protein,CD4 cel,MC4R – 37 kDa seven transmembrane G-protein coupled receptor,neurons,hypothalamus,brain,Adt2p – 34 kDa six transmembrane protein,yeast mitochondrial inner membrane,reconstitution,crystallization for X-ray diffraction,NMR experiments
Full Text Link:
Related URLs: http://www.micr ... ontent/5/S1/P38 (Publisher URL)
PURE Output Type: Article
Published Date: 2006-12
Authors: Jamshad, Mohammed
Darby, Richard A. J.
Grgic, Ljuban
Bill, Roslyn M. ( 0000-0003-1331-0852)

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