Novel Chemical Biological Methods for Detection of Enzyme Activity in Inflammation

Bioorthogonal coupling of small organic molecules to protein-bound functional groups is a long-standing challenge with many implications for cell biology. To further address this challenge we developed the oxidative Heck reaction as a novel strategy for bioorthogonal coupling of arylboronic acids to protein-bound alkenes¹. Further development of this reaction led to the identification of EDTA-Pd(II) as a novel catalyst for the oxidative Heck reaction on protein-bound alkenes, which can be employed in fully aqueous reaction conditions². This reaction was employed to monitor histone lysine acylation in vitro after metabolic incorporation of olefinic carboxylates as chemical reporters². Another successful application was found by application of the oxidative Heck reaction in a two-step activity-based protein profiling (ABPP) approach for lipoxygenase activity in biological samples, which is a class of enzymes that has not been addressed by ABPP so far³. Altogether these studies demonstrate that chemistry-based methods enable the investigation of enzyme activity in the context of inflammation.

[1] Ourailidou ME, Dockerty P, Witte M, Poelarends GJ, Dekker FJ. Metabolic alkene labeling and in vitro detection of histone acylation via the aqueous oxidative Heck reaction. Org. Biomol. Chem. 2015, 13(12), 3648-3653.

[2] Ourailidou ME, van der Meer JY, Baas BJ, Jeronimus-Stratingh M, Gottumukkala AL, Poelarends GJ, Minnaard AJ, Dekker FJ. Aqueous oxidative Heck reaction as a protein-labeling strategy. Chembiochem. 2014, 15(2), 209-212.

[3] Eleftheriadis N, Thee SA, Zwinderman MR, Leus NG, Dekker FJ. Activity-Based Probes for 15-Lipoxygenase-1. Angew Chem Int Ed Engl. 2016, 55(40), 12300-12305.