A toolbox of diverse arginine N-glycosylated peptides and specific antibodies

Among posttranslational modifications, atypical arginine N-glycosylation has drawn increasing interest due to its fundamental role in various cellular procedures and signaling pathways. The efficient synthesis of arginine N-glycosylated substrates, as well as the generation of specific antibodies, remains challenging. This work describes the efficient synthesis of diverse arginine N-glycosylated peptides, in a process termed silver-promoted solid -phase glycosylation (SGG). There are two key features of the SSG strategy: (i) robust synthesis of gram-scale S-alkyl-isothiourea glycosyl donors facilitates the subsequent SSG procedure and (ii) the simultaneous introduction of both the side-chain sugar motif and arginine residue. Notably, our findings, combined with our previous results, provide a toolbox containing diverse S-alkyl-isothiourea glycosyl donors (glucose, galactose, mannose, ribose, xylose, lactose and maltose) as well as the corresponding Arg N-glycosylated peptides. In addition, our toolbox is shown to help investigate specific antibodies and identify multiple potent and precise biochemical tools for exploring arginine N-glycosylation.

Automated summary

Among posttranslational modifications, atypical arginine N-glycosylation plays a fundamental role in cellular processes and signaling pathways. This work presents a silver-promoted solid-phase glycosylation (SGG) method for efficient synthesis of diverse arginine N-glycosylated peptides. The SSG strategy involves robust synthesis of gram-scale S-alkyl-isothiourea glycosyl donors and simultaneous introduction of side-chain sugar motif and arginine residue. The resulting toolbox facilitates the investigation of specific antibodies and provides potent biochemical tools for exploring arginine N-glycosylation.