title: 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.
abstract: 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.
authors.affiliation: Université de Lille, Sciences et Technologies, Unité de Glycobiologie Structurale et Fonctionnelle (UMR CNRS 8576), 59000, Lille, France.|CNRS, UMR 8576, 59000, Lille, France.
authors.forename: Haoling
authors.initials: H
authors.lastname: Qi
authors.affiliation: Université de Lille, Sciences et Technologies, Unité de Glycobiologie Structurale et Fonctionnelle (UMR CNRS 8576), 59000, Lille, France.|CNRS, UMR 8576, 59000, Lille, France.
authors.forename: Clément
authors.initials: C
authors.lastname: Despres
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA, 02115, USA.
authors.forename: Sudhakaran
authors.initials: S
authors.lastname: Prabakaran
authors.affiliation: Université de Lille, Sciences et Technologies, Unité de Glycobiologie Structurale et Fonctionnelle (UMR CNRS 8576), 59000, Lille, France.|CNRS, UMR 8576, 59000, Lille, France.
authors.forename: François-Xavier
authors.initials: FX
authors.lastname: Cantrelle
authors.affiliation: INSERM, Paris-Saclay University, UMR1195, Le Kremlin Bicêtre, France.
authors.forename: Béatrice
authors.initials: B
authors.lastname: Chambraud
authors.affiliation: Department of Systems Biology, Harvard Medical School, Boston, MA, 02115, USA.
authors.forename: Jeremy
authors.initials: J
authors.lastname: Gunawardena
authors.affiliation: Université de Lille, Sciences et Technologies, Unité de Glycobiologie Structurale et Fonctionnelle (UMR CNRS 8576), 59000, Lille, France.|CNRS, UMR 8576, 59000, Lille, France.
authors.forename: Guy
authors.initials: G
authors.lastname: Lippens
authors.affiliation: Université de Lille, Sciences et Technologies, Unité de Glycobiologie Structurale et Fonctionnelle (UMR CNRS 8576), 59000, Lille, France. caroline.smet@univ-lille1.fr.|CNRS, UMR 8576, 59000, Lille, France. caroline.smet@univ-lille1.fr.
authors.forename: Caroline
authors.initials: C
authors.lastname: Smet-Nocca
authors.affiliation: UMR8576 CNRS-Université de Lille, Sciences et Technologies, Résonance Magnétique Nucléaire (RMN), IRI Building (CNRS), Campus CNRS Parc de la Haute Borne50, avenue de Halley, Villeneuve d'Ascq, BP70478, 59658, Lille, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Landrieu
mesh_terms: D000107:Acetylation; D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism; D000818:Animals; D050882:CREB-Binding Protein / Q000235:genetics / Q000378:metabolism*; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010766:Phosphorylation; D011499:Protein Processing, Post-Translational / Q000235:genetics / Q000502:physiology*; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016875:tau Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D011994:Recombinant Proteins; D016875:tau Proteins; D050882:CREB-Binding Protein; D019950:Mitogen-Activated Protein Kinase 1
keywords: Acetylation; Acetyltransferase; Creb-binding protein; ERK kinase; NMR spectroscopy; Phosphorylation; Recombinant proteins