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

Angew Chem Weinheim Bergstr Ger. 2024 Jan 22;136(4):e202313317. doi: 10.1002/ange.202313317. Epub 2023 Nov 14.

Abstract

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 α-configured N(τ)- and N(π)-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(τ)‐ and N(π)‐ADP‐ribosylated histidine. The N‐glycosidic linkage proved stable to the treatment with aqueous acid and base and resistant to (ADP‐ribosyl)hydrolases.

Keywords: ADP-Ribosylation; Glycosylation; Histidine; Peptides; Solid-Phase Synthesis.