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.
Algal autolysate medium to label proteins for NMR in mammalian cells / Fuccio, Carmelo; Luchinat, Enrico; Barbieri, Letizia; Neri, Sara; Fragai, Marco. - In: JOURNAL OF BIOMOLECULAR NMR. - ISSN 0925-2738. - STAMPA. - 64:(2016), pp. 275-280. [10.1007/s10858-016-0026-0]
Algal autolysate medium to label proteins for NMR in mammalian cells
FUCCIO, CARMELO;LUCHINAT, ENRICO;BARBIERI, LETIZIA;NERI, SARA;FRAGAI, MARCO
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.File | Dimensione | Formato | |
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