Tau Protein pp 179-213 | Cite as

The Study of Posttranslational Modifications of Tau Protein by Nuclear Magnetic Resonance Spectroscopy: Phosphorylation of Tau Protein by ERK2 Recombinant Kinase and Rat Brain Extract, and Acetylation by Recombinant Creb-Binding Protein

  • Haoling Qi
  • Clément Despres
  • Sudhakaran Prabakaran
  • François-Xavier Cantrelle
  • Béatrice Chambraud
  • Jeremy Gunawardena
  • Guy Lippens
  • Caroline Smet-NoccaEmail author
  • Isabelle Landrieu
Part of the Methods in Molecular Biology book series (MIMB, volume 1523)


Nuclear magnetic resonance (NMR) spectroscopy can be used as an analytical tool to investigate posttranslational modifications of protein. NMR is a valuable tool to map the interaction regions of protein partners. Here, we present protocols that have been developed in the course of our studies of the neuronal Tau protein. Tau is found aggregated in the neurons of Alzheimer’s disease patients. Development of the disease is accompanied by increased, abnormal phosphorylation and acetylation of Tau. We have used NMR to investigate how these posttranslational modifications of Tau affect the interactions with its partners. We present here detailed protocols of in vitro phosphorylation of Tau by recombinant kinase, ERK2, or kinase activity of rat brain extracts, and acetylation by recombinant Creb-binding protein (CBP) acetyltransferase. The analytical characterization of the modified Tau by NMR spectroscopy is additionally described.

Key words

Phosphorylation Acetylation ERK kinase Creb-binding protein Acetyltransferase NMR spectroscopy Recombinant proteins 



The NMR facilities were funded by the Région Nord, CNRS, Pasteur Institute of Lille, European Community (FEDER), French Research Ministry and the University of Sciences and Technologies of Lille I. We acknowledge support from the TGE RMN THC (FR-3050, France) and the Research Federation FRABio (Univ. Lille, CNRS, FR 3688, FRABio, “Structural & Functional Biochemistry of Biomolecular Assemblies”) for providing the scientific and technical environment. This study was supported by a grant from the LabEx (Laboratory of Excellence) DISTALZ (Development of Innovative Strategies for a Transdisciplinary approach to Alzheimer’s disease), and in part by the French government funding agency Agence Nationale de la Recherche TAF. S.P. and J.G. were partially supported on NIH R01-GM08157.


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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Haoling Qi
    • 1
    • 2
  • Clément Despres
    • 1
    • 2
  • Sudhakaran Prabakaran
    • 3
  • François-Xavier Cantrelle
    • 1
    • 2
  • Béatrice Chambraud
    • 4
  • Jeremy Gunawardena
    • 3
  • Guy Lippens
    • 1
    • 2
  • Caroline Smet-Nocca
    • 1
    • 2
    Email author
  • Isabelle Landrieu
    • 5
  1. 1.Université de Lille, Sciences et Technologies, Unité de Glycobiologie Structurale et Fonctionnelle (UMR CNRS 8576)LilleFrance
  2. 2.CNRS, UMR 8576LilleFrance
  3. 3.Department of Systems BiologyHarvard Medical SchoolBostonUSA
  4. 4.INSERM, Paris-Saclay University, UMR1195Le Kremlin BicêtreFrance
  5. 5.UMR8576 CNRS-Université de Lille, Sciences et Technologies, Résonance Magnétique Nucléaire (RMN)LilleFrance

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