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Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins
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Journal JoVE Biochimie
Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins

Nuclear Magnetic Resonance Spectroscopy for the Identification of Multiple Phosphorylations of Intrinsically Disordered Proteins

DOI:
10.3791/55001-v

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12:47 min

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December 27, 2016

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, , , , , , , , , , , ,
1UMR8576, CNRS,Lille University, 2UMR-S1172, INSERM CNRS,Lille University

Chapitres

  • 00:05Titre
  • 01:15Production of 15N, 13C-Tau
  • 04:18Purification of 15N, 13C-Tau
  • 07:38Acquisition of NMR Spectra
  • 10:28Results: Evaluation of the Purification Procedure of the Recombinant Protein and NMR Spectrum
  • 11:35Conclusion

Summary

Traduction automatique

Traduction automatique

We describe here a method to identify multiple phosphorylations of an intrinsically disordered protein by Nuclear Magnetic Resonance Spectroscopy (NMR), using Tau protein as a case study. Recombinant Tau is isotopically enriched and modified in vitro by a kinase prior to data acquisition and analysis.

Tags

Nuclear Magnetic Resonance SpectroscopyIntrinsically Disordered ProteinsTau ProteinPhosphorylationMolecular RegulationDiseaseProtein InteractionProtein StructureProtein FunctionAPtc VirusTherapyProtein InhibitorsBL21 CellsPlasmid DNALuria Bertani MediumAmpicillinM9 MediumIsotope LabelingRecombinant PlasmidOptical Density

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