Methyltetrazine as a small live-cell compatible bioorthogonal handle for imaging enzyme activities in situ

RSC Chemical Biology, vol. 3, pp. 1325–1330

Abstract

Bioorthogonal chemistry combines well with activity-based protein profiling, as it allows for the introduction of detection tags without significantly influencing the physiochemical and biological functions of the probe. In this work, we introduced methyltetrazinylalanine (MeTz-Ala), a close mimic of phenylalanine, into a dipeptide fluoromethylketone cysteine protease inhibitor. Following covalent and irreversible inhibition, the tetrazine allows vizualisation of the captured cathepsin activity by means of inverse electron demand Diels Alder ligation in cell lysates and live cells, demonstrating that tetrazines can be used as live cell compatible, minimal bioorthogonal tags in activity-based protein profiling.

Authors 9

1. Diana Torres‐García Aachen Leiden Institute of Chemistry, The Institute for Chemical Immunology

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

2. Merel A. T. van de Plassche Aachen Leiden Institute of Chemistry, The Institute for Chemical Immunology

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

3. Emma van Boven Aachen Leiden Institute of Chemistry, The Institute for Chemical Immunology

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

4. Tyrza van Leeuwen Aachen Leiden Institute of Chemistry, The Institute for Chemical Immunology

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

5. Mirjam G. J. Groenewold Aachen Leiden Institute of Chemistry, The Institute for Chemical Immunology

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

6. Alexi J. C. Sarris Aachen Leiden Institute of Chemistry, The Institute for Chemical Immunology

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

7. Luuk Klein Aachen Leiden Institute of Chemistry, The Institute for Chemical Immunology

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

8. Herman S. Overkleeft corresponding Aachen Leiden Institute of Chemistry, The Institute for Chemical Immunology

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

9. Sander I. van Kasteren corresponding Aachen Leiden Institute of Chemistry, The Institute for Chemical Immunology

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry and The Institute for Chemical Immunology, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands