Tau Protein pp 215-235 | Cite as

Tag-Free Semi-Synthesis of the Tau Protein

  • Oliver ReimannEmail author
  • Caroline Smet-Nocca
  • Christian P. R. Hackenberger
Part of the Methods in Molecular Biology book series (MIMB, volume 1523)


Expressed protein ligation (EPL) is a valuable tool to study site-specific functionalities on proteins such as posttranslational modifications. The purification of such ligation products from EPL mixtures can be cumbersome due to a small size difference between the expressed protein portion and the desired ligated protein. Therefore, affinity tags are often required, which remain on the protein after purification. Herein, we present an efficient protocol to install a photocleavable biotin building block on synthetic C-terminal tau[390–441] and describe its use for purification of full-length semi-synthetic tau[1–441].

Key words

Expressed protein ligation Photocleavable biotin Semi-synthesis Affinity purification Traceless linker 


  1. 1.
    Goedert M (1993) Tau-protein and the neurofibrillary pathology of Alzheimers-disease. Trends Neurosci 16:460–465CrossRefPubMedGoogle Scholar
  2. 2.
    Buee L, Bussiere T, Buee-Scherrer V, Delacourte A, Hof PR (2000) Tau protein isoforms, phosphorylation and role in neurodegenerative disorders. Brain Res Rev 33:95–130CrossRefPubMedGoogle Scholar
  3. 3.
    Trojanowski JQ, Lee VMY (1995) Phosphorylation of paired helical filament tau in Alzheimer’s disease neurofibrillary lesions: focusing on phosphatases. FASEB J 9:1570–1576PubMedGoogle Scholar
  4. 4.
    Patrick GN, Zukerberg L, Nikolic M, de la Monte S, Dikkes P, Tsai LH (1999) Conversion of p35 to p25 deregulates Cdk5 activity and promotes neurodegeneration. Nature 402:615–622CrossRefPubMedGoogle Scholar
  5. 5.
    Lindwall G, Cole RD (1984) Phosphorylation affects the ability of tau-protein to promote microtubule assembly. J Biol Chem 259:5301–5305PubMedGoogle Scholar
  6. 6.
    Alonso AD, Zaidi T, Grundkeiqbal I, Iqbal K (1994) Role of abnormally phosphorylated tan in the breakdown of microtubules in Alzheimer-disease. Proc Natl Acad Sci U S A 91:5562–5566CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Abraha A, Ghoshal N, Gamblin TC, Cryns V, Berry RW, Kuret J, Binder LI (2000) C-terminal inhibition of tau assembly in vitro and in Alzheimer’s disease. J Cell Sci 113:3737–3745PubMedGoogle Scholar
  8. 8.
    Dawson PE, Muir TW, Clark-Lewis I, Kent SB (1994) Synthesis of proteins by native chemical ligation. Science 266:776–779CrossRefPubMedGoogle Scholar
  9. 9.
    Muir TW, Sondhi D, Cole PA (1998) Expressed protein ligation: a general method for protein engineering. Proc Natl Acad Sci U S A 95:6705–6710CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Hackenberger CP, Schwarzer D (2008) Chemoselective ligation and modification strategies for peptides and proteins. Angew Chem Int Ed Engl 47:10030–10074CrossRefPubMedGoogle Scholar
  11. 11.
    Broncel M, Krause E, Schwarzer D, Hackenberger CP (2012) The Alzheimer’s disease related tau protein as a new target for chemical protein engineering. Chem Eur J 18:2488–2492CrossRefPubMedGoogle Scholar
  12. 12.
    Reimann O, Smet-Nocca C, Hackenberger CPR (2015) Traceless purification and desulfurization of tau protein ligation products. Angew Chem Int Ed 54:306–310CrossRefGoogle Scholar
  13. 13.
    Reimann O, Glanz M, Hackenberger CP (2015) Native chemical ligation between asparagine and valine: application and limitations for the synthesis of tri-phosphorylated C-terminal tau. Bioorg Med Chem 23:2890–2894CrossRefPubMedGoogle Scholar
  14. 14.
    Schwagerus S, Reimann O, Despres C, Smet-Nocca C, Hackenberger CP (2016) Semisynthesis of a tag-free O-GlcNAcylated tau protein by sequential hemoselective ligation. J Pept Sci 22:327–333CrossRefPubMedGoogle Scholar
  15. 15.
    Bindman N, Merkx R, Koehler R, Herrman N, van der Donk WA (2010) Photochemical cleavage of leader peptides. Chem Commun (Camb) 46:8935–8937CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Oliver Reimann
    • 1
    • 2
    Email author
  • Caroline Smet-Nocca
    • 3
    • 4
  • Christian P. R. Hackenberger
    • 1
    • 5
  1. 1.Leibniz-Institute for Molecular Pharmacology (FMP)BerlinGermany
  2. 2.Freie Universitaet Berlin, Institute for Chemistry und BiochemistryBerlinGermany
  3. 3.Univ. Lille, CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et FonctionnelleLilleFrance
  4. 4.Research Federation FraBio 3688Villeneuve d’AscqFrance
  5. 5.Department of ChemistryHumboldt Universitaet zu BerlinBerlinGermany

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