Abstract
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.
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Acknowledgments
This work has been supported by MIUR—Project: BIOLABEL “Valorizzazione della biomassa algale per la marcatura isotopica delle biomolecole”.
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Carmelo Fuccio and Enrico Luchinat have contributed equally to this work.
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Fuccio, C., Luchinat, E., Barbieri, L. et al. Algal autolysate medium to label proteins for NMR in mammalian cells. J Biomol NMR 64, 275–280 (2016). https://doi.org/10.1007/s10858-016-0026-0
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DOI: https://doi.org/10.1007/s10858-016-0026-0