Disulphide-mediated site-directed modification of proteins

Abstract

<p>Site-directed addition of a single thiols handle to proteins by means of temporary disulphide rebridging of solvent exposed disulphides is obtained with a new labelling reagent.</p>

DNAzymes for amine and peptide lysine acylation

“…74 Expanding the substrate tolerance of such DNAzymes from peptides to proteins is a further challenge, but worth undertaking considering the difficulty inherent to achieving nonenzymatic site-selective Lys modification of native proteins. [59][60][61][62][63][64][65] We are currently pursuing such experiments.…”

“…25,26 One such reaction is lysine (Lys) acylation, where Lys acetylation is critical for histones and in other contexts, [27][28][29][30] and many longer-chain Lys acylation PTMs [31][32][33] such as malonylation, 34,35 succinylation, 34,36 and glutarylation 37,38 have been discovered yet are poorly understood. 39 As an alternative to approaches that include introduction of Lys analogues, [40][41][42][43] nonsense codon suppression, [44][45][46][47][48][49] bottom-up ligation-based assembly strategies, [50][51][52] or enzymatic methods that typically require creation of a non-native protein by insertion of a specific target sequence, [53][54][55][56][57][58] DNAzymes are promising for top-down introduction of Lys acylation PTMs onto intact native proteins, [59][60][61][62][63][64][65] but only if DNAzymes can be identified with the fundamental catalytic ability of Lys acylation.…”

Proximity‐driven, Regioselective Chemical Modification of Peptides and Proteins

“…Finally, Gothelf, Clo and co-workers have recently reported a method to generate "pseudo-cysteine mutants", in which Lys residues can be regioselectively modified with a masked thiol reagent using a nearby disulfide to provide proximity-based selectivity. [96] First, a disulfide within the protein of interest is reduced and intercalated by a bis-pyridyl disulfide reagent. Proximity-based imine formation to a nearby Lys residue is covalently quenched by reductive amination.…”

A Reagent for Amine‐Directed Conjugation to IgG1 Antibodies

“…Targeting more universal sites such as metal binding sites or disulfides, allows for the utilization of the same reagent or strategy on different proteins [15] . Recently, we, and Forte et al., published two related approaches for disulfide directed labeling of proteins [20, 21] . The metal chelator tris‐nitrilotriacetic acid has been used in different approaches for targeting metal‐binding sites [22, 23] .…”