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Triple Bioorthogonal Ligation Strategy for Simultaneous Labeling of Multiple Enzymatic Activities

Authors

  • Lianne I. Willems,

    1. Leiden Institute of Chemistry and Netherlands Proteomics Centre, Gorlaeus Laboratories, Einsteinweg 55, 2333 CC Leiden (The Netherlands)
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  • Nan Li,

    1. Leiden Institute of Chemistry and Netherlands Proteomics Centre, Gorlaeus Laboratories, Einsteinweg 55, 2333 CC Leiden (The Netherlands)
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  • Dr. Bogdan I. Florea,

    1. Leiden Institute of Chemistry and Netherlands Proteomics Centre, Gorlaeus Laboratories, Einsteinweg 55, 2333 CC Leiden (The Netherlands)
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  • Mark Ruben,

    1. Leiden Institute of Chemistry and Netherlands Proteomics Centre, Gorlaeus Laboratories, Einsteinweg 55, 2333 CC Leiden (The Netherlands)
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  • Prof. Gijsbert A. van der Marel,

    1. Leiden Institute of Chemistry and Netherlands Proteomics Centre, Gorlaeus Laboratories, Einsteinweg 55, 2333 CC Leiden (The Netherlands)
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  • Prof. Herman S. Overkleeft

    Corresponding author
    1. Leiden Institute of Chemistry and Netherlands Proteomics Centre, Gorlaeus Laboratories, Einsteinweg 55, 2333 CC Leiden (The Netherlands)
    • Leiden Institute of Chemistry and Netherlands Proteomics Centre, Gorlaeus Laboratories, Einsteinweg 55, 2333 CC Leiden (The Netherlands)
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  • This research was financially supported by the Netherlands Organisation for Scientific Research (NWO) and the Netherlands Genomics Initiative (NGI).

Abstract

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Three at the same time: A ligation strategy combining tetrazine–norbornene cycloaddition, Staudinger–Bertozzi ligation, and copper(I)-catalyzed click reaction was used to label the three catalytic activities of the proteasome simultaneously in a single experiment. The orthogonality of the three ligation reactions enables selective monitoring of multiple targets at the same time in complex biological samples.

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