Algal autolysate medium to label proteins for NMR in mammalian cells

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...

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Published inJournal of biomolecular NMR Vol. 64; no. 4; pp. 275 - 280
Main Authors Fuccio, Carmelo, Luchinat, Enrico, Barbieri, Letizia, Neri, Sara, Fragai, Marco
Format Journal Article
LanguageEnglish
Published Dordrecht Springer Netherlands 01.04.2016
Springer Nature B.V
Subjects
Animals
Biochemistry
Biological and Medical Physics
Biophysics
Cell Line
Cells, Cultured
Cyanobacteria - metabolism
Humans
Isotope Labeling
Mammals
Nuclear Magnetic Resonance, Biomolecular
Physics
Physics and Astronomy
Proteins
Proteins - chemistry
Proteins - genetics
Proteins - metabolism
Spectroscopy/Spectrometry
Spirulina platensis
Staining and Labeling
Cell culture
Protein expression
In-cell NMR
Mammalian cells
Isotopic labeling
Spirulina platensis
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Summary: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 15 N-enriched S. platensis algal-derived medium, and high quality in-cell NMR spectra were obtained.
Bibliography:ObjectType-Article-1
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ISSN:0925-2738
1573-5001
DOI:10.1007/s10858-016-0026-0