Should I Stay or Should I Go—Leveraging an Overlooked Feature of Click‐to‐Release Chemistry for Affinity Labeling
5 affiliations
- 1MTA – HUN‐REN TTK Lendület “Momentum” Chemical Biology Research Group Institute of Organic Chemistry HUN‐REN Research Centre For Natural Sciences Budapest Hungary
- 2Hevesy György PhD School of Chemistry Eötvös Loránd University Budapest Hungary
- 3MS Metabolomic Research Laboratory Centre For Structural Science HUN‐REN Research Centre For Natural Sciences Budapest Hungary
- 4Medicinal Chemistry Research Group Institute of Organic Chemistry HUN‐REN Research Centre For Natural Sciences Budapest Hungary
- 5Institute of Chemistry Eötvös Loránd University Budapest Hungary
Abstract
Click‐to‐release (C2R) chemistry is widely used for controlled payload liberation, yet the reactivity of its transient intermediates has remained largely unexplored. Here we show that an intermediate formed during the reaction of trans‐cyclooctene (TCO)‐appended protein‐targeting ligands with release tetrazines (rTzs), previously regarded as a byproduct rapidly quenched by water, can be exploited as an electrophile for ligand‐directed covalent protein labeling. The bioorthogonally generated dihydropyridazine methide (DHP‐methide) enables affinity‐directed protein labeling, resulting primarily in capture at surface‐exposed histidine residues proximal to the site of electrophile generation and revealing a histidine‐biased covalent modification pathway. Incorporation of functional handles into the rTz platform enables modular reporter installation via an orthogonal conjugation step. A fluorogenic leaving‐group design further allows direct optical reporting of the electrophile‐generating elimination event and real‐time monitoring of DHP‐methide formation. Selective labeling is retained in cell lysate and can be translated to a cellular context, as demonstrated by confocal imaging of surface‐expressed targets. These findings establish a bioorthogonally gated strategy for affinity labeling and reveal a previously unrecognized functional dimension of rTz–TCO click‐to‐release chemistry.
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BibTeX
@article{kern2026should,
title = {{Should I Stay or Should I Go—Leveraging an Overlooked Feature of Click‐to‐Release Chemistry for Affinity Labeling}},
author = {Dóra Kern and Krisztina Németh and Tímea Imre and József Simon and Kitti Meggyes and Gergő Bárdos and László Petri and Márton Bojtár and Attila Kormos and Péter Kele},
journal = {Angewandte Chemie (International Ed. in English)},
year = {2026},
month = aug,
doi = {10.1002/anie.5715165},
url = {https://doi.org/10.1002/anie.5715165},
note = {CC BY-NC 4.0},
}
RIS
TY - JOUR
TI - Should I Stay or Should I Go—Leveraging an Overlooked Feature of Click‐to‐Release Chemistry for Affinity Labeling
AU - Dóra Kern
AU - Krisztina Németh
AU - Tímea Imre
AU - József Simon
AU - Kitti Meggyes
AU - Gergő Bárdos
AU - László Petri
AU - Márton Bojtár
AU - Attila Kormos
AU - Péter Kele
PY - 2026
DA - 2026/08/26
JO - Angewandte Chemie (International Ed. in English)
DO - 10.1002/anie.5715165
UR - https://doi.org/10.1002/anie.5715165
AB - Click‐to‐release (C2R) chemistry is widely used for controlled payload liberation, yet the reactivity of its transient intermediates has remained largely unexplored. Here we show that an intermediate formed during the reaction of trans‐cyclooctene (TCO)‐appended protein‐targeting ligands with release tetrazines (rTzs), previously regarded as a byproduct rapidly quenched by water, can be exploited as an electrophile for ligand‐directed covalent protein labeling. The bioorthogonally generated dihydropyridazine methide (DHP‐methide) enables affinity‐directed protein labeling, resulting primarily in capture at surface‐exposed histidine residues proximal to the site of electrophile generation and revealing a histidine‐biased covalent modification pathway. Incorporation of functional handles into the rTz platform enables modular reporter installation via an orthogonal conjugation step. A fluorogenic leaving‐group design further allows direct optical reporting of the electrophile‐generating elimination event and real‐time monitoring of DHP‐methide formation. Selective labeling is retained in cell lysate and can be translated to a cellular context, as demonstrated by confocal imaging of surface‐expressed targets. These findings establish a bioorthogonally gated strategy for affinity labeling and reveal a previously unrecognized functional dimension of rTz–TCO click‐to‐release chemistry.
SN - 1433-7851
SN - 1521-3773
ER -