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

Fingerprint

Yeast
Membrane Proteins
Yeasts
CD63 Antigen
Saccharomyces cerevisiae
Proteins
Mitochondrial Membranes
Cell growth
Structure-Activity Relationship
Post Translational Protein Processing
Crystallization
Eukaryota
Viruses
X-Ray Diffraction
Hepacivirus
Adenosine Diphosphate
Hypothalamus
Neurons
Lymphoma
Brain

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

Cite this

Jamshad, Mohammed ; Darby, Richard A. J. ; Grgic, Ljuban ; Bill, Roslyn M. / Production of membrane proteins in yeast. In: Biochemistry and Cell Biology. 2006 ; Vol. 84, No. 6. pp. 1069.
@article{9058ee0f7c4c49cf81f552a6fb008461,
title = "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.",
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",
author = "Mohammed Jamshad and Darby, {Richard A. J.} and Ljuban Grgic and Bill, {Roslyn M.}",
note = "{\circledC} Darby et al; licensee BioMed Central Ltd. 2006",
year = "2006",
month = "12",
doi = "10.1186/1475-2859-5-S1-P38",
language = "English",
volume = "84",
pages = "1069",
journal = "Biochemistry and Cell Biology",
issn = "0829-8211",
publisher = "National Research Council of Canada",
number = "6",

}

Production of membrane proteins in yeast. / Jamshad, Mohammed; Darby, Richard A. J.; Grgic, Ljuban; Bill, Roslyn M.

In: Biochemistry and Cell Biology, Vol. 84, No. 6, 12.2006, p. 1069.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Production of membrane proteins in yeast

AU - Jamshad, Mohammed

AU - Darby, Richard A. J.

AU - Grgic, Ljuban

AU - Bill, Roslyn M.

N1 - © Darby et al; licensee BioMed Central Ltd. 2006

PY - 2006/12

Y1 - 2006/12

N2 - 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.

AB - 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.

KW - yeast

KW - membrane proteins

KW - eukaryote-like post-translational modifications

KW - S. cerevisiae

KW - fully-sequenced genom

KW - P. pastoris

KW - cell densities

KW - lymphoma cell growth

KW - viral receptor

KW - Hepatitis C-Virus

KW - CD82 – 30 kDa tetraspanin protein

KW - CD4 cel

KW - MC4R – 37 kDa seven transmembrane G-protein coupled receptor

KW - neurons

KW - hypothalamus

KW - brain

KW - Adt2p – 34 kDa six transmembrane protein

KW - yeast mitochondrial inner membrane

KW - reconstitution

KW - crystallization for X-ray diffraction

KW - NMR experiments

UR - http://www.microbialcellfactories.com/content/5/S1/P38

U2 - 10.1186/1475-2859-5-S1-P38

DO - 10.1186/1475-2859-5-S1-P38

M3 - Article

VL - 84

SP - 1069

JO - Biochemistry and Cell Biology

JF - Biochemistry and Cell Biology

SN - 0829-8211

IS - 6

ER -