Production of isotope-labeled proteins in insect cells for NMR
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Baculovirus-infected insect cells have become a powerful tool to express recombinant proteins for structural and functional studies by NMR spectroscopy. This article provides an introduction into the insect cell/baculovirus expression system and its use for the production of recombinant isotope-labeled proteins. We discuss recent advances in inexpensive isotope-labeling methods using labeled algal or yeast extracts as the amino acid source and give examples of advanced NMR applications for proteins, which have become accessible by this eukaryotic expression host.
KeywordsBaculovirus Yeast extract Algal extract Deuteration GPCR Abelson kinase
This work was supported by the Swiss National Science Foundation (Grants 31-149927 and 31-173089 to S.G.), the Swiss Cancer League (Grant KFS-3603-02-2015 to S.G.) and the Biozentrum Basel International PhD Program (Fellowship of Excellence to A.G.). We gratefully acknowledge the Biozentrum’s proteomics and biophysics core facilities for expert mass spectrometric analysis as well as Drs. Alexander Schmidt, Andre Strauss and Binesh Shrestha for stimulating discussions.
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