Algal autolysate medium to label proteins for NMR in mammalian cells
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In-cell NMR provides structural and functional information on proteins directly inside living cells. At present, the high costs of the labeled media for mammalian cells represent a limiting factor for the development of this methodology. Here we report a protocol to prepare a homemade growth medium from Spirulina platensis autolysate, suitable to express uniformly labeled proteins inside mammalian cells at a reduced cost-per-sample. The human proteins SOD1 and Mia40 were overexpressed in human cells grown in 15N-enriched S. platensis algal-derived medium, and high quality in-cell NMR spectra were obtained.
KeywordsIn-cell NMR Cell culture Mammalian cells Spirulina platensis Protein expression Isotopic labeling
This work has been supported by MIUR—Project: BIOLABEL “Valorizzazione della biomassa algale per la marcatura isotopica delle biomolecole”.
- Freshney R (1987) Culture of animal cells: a manual of basic technique. Alan R. Liss Inc, New YorkGoogle Scholar