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.
FUCCIO, C., LUCHINAT, E., BARBIERI, L., NERI, S., FRAGAI, M. (2016). Algal autolysate medium to label proteins for NMR in mammalian cells. JOURNAL OF BIOMOLECULAR NMR, 64, 275-280 [10.1007/s10858-016-0026-0].
Algal autolysate medium to label proteins for NMR in mammalian cells
LUCHINAT, ENRICO;
2016
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.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
