Solid-Phase Synthesis and Biological Evaluation of Peptides ADP-Ribosylated at Histidine

The transfer of an adenosine diphosphate (ADP) ribose moiety to a nucleophilic side chain by consumption of nicotinamide adenine dinucleotide is referred to as ADP-ribosylation, which allows for the spatiotemporal regulation of vital processes such as apoptosis and DNA repair. Recent mass-spectrometry based analyses of the "ADP-ribosylome" have identified histidine as ADP-ribose acceptor site. In order to study this modification, a fully synthetic strategy towards alpha-configured N(tau)- and N(pi)-ADP-ribosylated histidine-containing peptides has been developed. Ribofuranosylated histidine building blocks were obtained via Mukaiyama-type glycosylation and the building blocks were integrated into an ADP-ribosylome derived peptide sequence using fluorenylmethyloxycarbonyl (Fmoc)-based solid-phase peptide synthesis. On-resin installation of the ADP moiety was achieved using phosphoramidite chemistry, and global deprotection provided the desired ADP-ribosylated oligopeptides. The stability under various chemical conditions and resistance against (ADP-ribosyl) hydrolase-mediated degradation has been investigated to reveal that the constructs are stable under various chemical conditions and non-degradable by any of the known ADP-ribosylhydrolases. We report the preparation of ribofuranosylated Fmoc-histidine building blocks via a Mukaiyama-type glycosylation. These building blocks were used in solid-phase peptide synthesis, followed by on-resin pyrophosphate construction and deprotection to access peptides containing N(tau)- and N(pi)-ADP-ribosylated histidine. The N-glycosidic linkage proved stable to the treatment with aqueous acid and base and resistant to (ADP-ribosyl)hydrolases.image