Recently, enzymes thought to be protein lysine deacetylases, have been shown to target different types of acyl posttranslational modifications (PTMs) on lysine residues, including crotonyl, glutaryl, and myristoyl. In order to investigate the hypothesis that, yet unknown, proteins of biochemical importance may recognize these post-translational modifications, we have developed a novel set of affinity-based probes (ABPs).<br/>Our ABPs are decameric peptides based on fragments of proteins known to carry the novel acyl modifications. Additionally, the probes are equipped with an alkyne handle for click chemistry as well as a photo cross-linker for covalently trapping of putative interacting proteins. Because such ABPs are aimed at investigating enzymes that recognize or cleave these modifications, they are potential substrates for some of the target enzymes. This will cause cleavage of the PTMs in the probes during assays using active enzymes, thus providing unambiguous results. In order to circumvent this cleavage, we introduce thioamide and hydrazide motifs as acyl group analogs. These motifs are known to inhibit NAD<sup>+</sup>-dependent deacylase enzymes (sirtuins). We envision that these motifs will improve the structural integrity of our probes in cellular environments.

Affinity-based probes to investigate posttranslational modifications

256th National Meeting (2018)

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