Journal of Biomolecular NMR

, Volume 64, Issue 4, pp 275–280

Algal autolysate medium to label proteins for NMR in mammalian cells

  • Carmelo Fuccio
  • Enrico Luchinat
  • Letizia Barbieri
  • Sara Neri
  • Marco Fragai
Article

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.

Keywords

In-cell NMR Cell culture Mammalian cells Spirulina platensis Protein expression Isotopic labeling 

Supplementary material

10858_2016_26_MOESM1_ESM.docx (914 kb)
Supplementary material 1 (DOCX 913 kb)

Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Carmelo Fuccio
    • 1
  • Enrico Luchinat
    • 1
    • 2
  • Letizia Barbieri
    • 1
    • 3
  • Sara Neri
    • 3
  • Marco Fragai
    • 1
    • 4
  1. 1.Magnetic Resonance Center (CERM)University of FlorenceSesto Fiorentino, FlorenceItaly
  2. 2.Department of Biomedical, Clinical and Experimental SciencesUniversity of FlorenceFlorenceItaly
  3. 3.Giotto Biotech S.R.L.Sesto Fiorentino, FlorenceItaly
  4. 4.Department of Chemistry “Ugo Schiff”University of FlorenceSesto Fiorentino, FlorenceItaly

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