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Formation of Ubiquitin Dimers via Azide–Alkyne Click Reaction

  • Silvia Eger
  • Martin Scheffner
  • Andreas Marx
  • Marina RubiniEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 832)

Abstract

The conjugation of poly-ubiquitin chains is a widespread post-translational modification of proteins that plays a role in many different cellular processes. Notably, the biological function of the attached ubiquitin chain depends on which lysine residue is used for chain formation. Here, we report a method for the modular synthesis of site-specifically linked ubiquitin dimers, which is based on click reaction between two artificial amino acids. In this way, it is possible to synthesize all seven naturally occurring ubiquitin connectivities, thus giving access to all ubiquitin dimers. Furthermore, this method can be generally applied to link ubiquitin to any substrate protein or even to link any two proteins site specifically.

Key words

Ubiquitin chains Artificial amino acids Azidohomoalanine (Aha) Pyrrolysine analogue (Plk) Click reaction Cu(I)-catalyzed Huisgen azide–alkyne cycloaddition Triazole linkage 

Notes

Acknowledgements

We gratefully acknowledge funding by the Boehringer Ingelheim Fonds (S.E.), the EU-network of excellence RUBICON (M.S.) and the DFG (M.S., A.M.).

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Silvia Eger
    • 1
  • Martin Scheffner
    • 2
  • Andreas Marx
    • 1
  • Marina Rubini
    • 1
    Email author
  1. 1.Department of Chemistry, Konstanz Research School Chemical BiologyUniversity of KonstanzKonstanzGermany
  2. 2.Department of Biology, Konstanz Research School Chemical BiologyUniversity of KonstanzKonstanzGermany

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