AbstractADP‐ribosylation is a complex post‐translational modification involved in key physiological processes and associated with various health and disease states. The growing interest in ADP‐ribosylation necessitates straightforward and efficient synthetic methods for the preparation of ADP‐ribosylated peptides/proteins. In this study, we report a facile reaction between nicotinamide adenine dinucleotide (NAD+) and alcohols promoted by a combination of ionic liquids, yielding up to 94 % with α : β ratios ranging from 88 : 12 to 99 : 1 and a switchable configuration selectivity. This method significantly simplifies the production of ADP‐ribosylated peptides and proteins, enabling diverse applications. It allows detailed investigation of side‐chain structure–activity relationships, facilitates two‐step clickable conjugation of ADP‐ribosyl groups to proteins, and, for the first time, enables non‐enzymatic synthesis of well‐defined ADP‐ribosylated peptides/proteins from natural counterparts and NAD+.
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