METHODS AND KITS FOR DETERMINING DEACYLASE ACTIVITY

Methods and kits for determining deacylase activity on an acylated lysine residue of a target polypeptide are disclosed. The methods disclosed comprise: providing one or more bacterial cells comprising: (i) an orthogonal pair of tRNA synthetase and tRNA; and/or (ii) a polynucleotide sequence encoding a target polypeptide, wherein said polynucleotide sequence comprises at least one codon that is specifically recognized by said tRNA; and (iii) at least one mammalian deacetylase; incubating said bacterial cell or extract thereof in the presence of Nɛ-acetyl lysine, wherein said tRNA is aminoacylated with Nɛ-modified lysine and incorporates the Nɛ-modified lysine into said target polypeptide, thereby expressing said target polypeptide comprising an Nɛ-modified lysine residue; determining the acylation levels of said target polypeptide; and comparing said acylation levels to a control; thereby determining deacylase activity.