Steric-Free Bioorthogonal Labeling of Acetylation Substrates Based on a Fluorine-Thiol Displacement Reaction (FTDR)

Abstract

We have developed a novel bioorthogonal reaction that can selectively displace fluorine substitutions alpha to amide bonds. This fluorine-thiol displacement reaction (FTDR) allows for fluorinated cofactors or precursors to be utilized as chemical reporters; hijacking acetyltransferase mediated acetylation both in vitro and in live cells, which cannot be achieved with azide- or alkyne-based chemical reporters. The fluorine labels can be further converted to biotin or fluorophore tags using FTDR, enabling the general detection and imaging of acetyl substrates. This strategy may lead to a steric-free labeling platform for substrate proteins, expanding our chemical toolbox for functional annotation of post-translational modifications (PTMs) in a systematic manner.

Identification and Profiling of Histone Acetyltransferase Substrates by Bioorthogonal Labeling

“…However, the catalyzing enzymes (HATs) for these identified targets were not elucidated, limiting the information provided by the acetylome identification. More recently, Wang and coworkers used fluoroacetate and fluoroacetyl-CoA in conjugation with reactive thiol agents to profile the whole cellular acetylome (Lyu et al, 2021).…”