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  Structural genomics on membrane proteins: comparison of more than 100 GPCRs in 3 expression systems

Lundstrom, K., Wagner, R., Reinhart, C., Desmyter, A., Cherouati, N., Magnin, T., et al. (2006). Structural genomics on membrane proteins: comparison of more than 100 GPCRs in 3 expression systems. Journal of Structural and Functional Genomics, 7(2), 77-91. doi:10.1007/s10969-006-9011-2.

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 Creators:
Lundstrom, Kenneth1, Author
Wagner, Renaud2, Author
Reinhart, Christoph3, Author              
Desmyter, Aline4, Author
Cherouati, Nadia2, Author
Magnin, Thierry2, Author
Zeder-Lutz, Gabrielle2, Author
Courtot, Melanie2, Author
Prual, Cécile3, Author              
André, Nicolas3, Author              
Hassaine, Gherici1, Author
Michel, Hartmut3, Author              
Cambillau, Christian4, Author
Pattus, Franc2, Author
Affiliations:
1Bioxtal, Chemin des Croisettes 22, Epalinges CH-1066, Switzerland., ou_persistent22              
2UMR7175 Department of Receptors and Membrane Proteins, Illkirch, France, ou_persistent22              
3Department of Molecular Membrane Biology, Max Planck Institute of Biophysics, Max Planck Society, ou_2068290              
4University of Marseille, Marseille, France, ou_persistent22              

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Free keywords: Expression vector, G protein-coupled receptors, Structural genomics
 Abstract: Production of recombinant receptors has been one of the major bottlenecks in structural biology on G protein-coupled receptors (GPCRs). The MePNet (Membrane Protein Network) was established to overexpress a large number of GPCRs in three major expression systems, based on Escherichia coli, Pichia pastoris and Semliki Forest virus (SFV) vectors. Evaluation by immunodetection demonstrated that 50% of a total of 103 GPCRs were expressed in bacterial inclusion bodies, 94% in yeast cell membranes and 95% in SFV-infected mammalian cells. The expression levels varied from low to high and the various GPCR families and subtypes were analyzed for their expressability in each expression system. More than 60% of the GPCRs were expressed at milligram levels or higher in one or several systems, compatible to structural biology applications. Functional activity was determined by binding assays in yeast and mammalian cells and the correlation between immunodetection and binding activity was analyzed.

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Language(s): eng - English
 Dates: 2006-04-192006-08-172006-11-192006-06
 Publication Status: Published in print
 Pages: 15
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1007/s10969-006-9011-2
PMID: 17120110
 Degree: -

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Title: Journal of Structural and Functional Genomics
  Other : J Struct Funct Genomics
Source Genre: Journal
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Publ. Info: Dordrecht : Kluwer Academic Publishers
Pages: - Volume / Issue: 7 (2) Sequence Number: - Start / End Page: 77 - 91 Identifier: ISSN: 1345-711X
CoNE: https://pure.mpg.de/cone/journals/resource/110989422336030