Production of membrane proteins in yeast

Mohammed Jamshad, Richard A. J. Darby, Ljuban Grgic, Roslyn M. Bill

Research output: Contribution to journalArticle

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.
Original languageEnglish
Pages (from-to)1069
Number of pages1
JournalBiochemistry and Cell Biology
Volume84
Issue number6
DOIs
Publication statusPublished - Dec 2006

Bibliographical note

© Darby et al; licensee BioMed Central Ltd. 2006

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

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