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. 2018 Jul 6;20(13):4140-4143.
doi: 10.1021/acs.orglett.8b01742. Epub 2018 Jun 27.

Synthetic α- and β-Ser-ADP-ribosylated Peptides Reveal α-Ser-ADPr as the Native Epimer

Jim Voorneveld  1 , Johannes G M Rack  2 , Ivan Ahel  2 , Herman S Overkleeft  1 , Gijsbert A van der Marel  1 , Dmitri V Filippov  1
Affiliations

Synthetic α- and β-Ser-ADP-ribosylated Peptides Reveal α-Ser-ADPr as the Native Epimer

Jim Voorneveld et al. Org Lett. .

Abstract

A solid-phase methodology to synthesize oligopeptides, specifically incorporating serine residues linked to ADP-ribose (ADPr), is presented. Through the synthesis of both α- and β-anomers of the phosphoribosylated Fmoc-Ser building block and their usage in our modified solid-phase peptide synthesis protocol, both α- and β-ADPr peptides from a naturally Ser-ADPr containing H2B sequence were obtained. With these, and by digestion studies using the human glycohydrolase, ARH3 (hARH3), compelling evidence is obtained that the α-Ser-ADPr linkage comprises the naturally occurring configuration.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Scheme 1
Scheme 1. Retrosynthetic Analysis for the Synthesis of Mono-ADP-ribosylated Peptides 1 and 2 and Essential Building Blocks
Scheme 2
Scheme 2. Synthesis of Orthogonally Protected α-Phosphoribosylated Serine
Scheme 3
Scheme 3. Synthesis of the β-Phosphoribosylated Serine Building Block
Figure 1
Figure 1
Analysis of the stereospecificity of hARH3. ADPr released in the ARH3 reaction was converted into AMP using hNUDT5 and subsequently measured using the AMP-Glo assay (Promega). Control reactions were carried out both in absence of hARH3 as well as using a catalytically inactive hARH3 mutant (D77N D788N). The data were normalized to reactions containing only hNUDT5 and represent triplicate measurement ± SD.
Scheme 4
Scheme 4. Mechanism of ADPr Elimination by NaOH
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