ADPr-Peptide Synthesis

  • Hans A. V. Kistemaker
  • Jim Voorneveld
  • Dmitri V. Filippov
Part of the Methods in Molecular Biology book series (MIMB, volume 1813)


Synthetic mono-ADPr-peptides are useful for structural, biochemical, and proteomics studies. We describe here a protocol for the preparation of mono-ADPr-peptides based on a fairly standard Fmoc-based solid-phase synthesis. Phosphoribosylated precursor building blocks are introduced into the peptide chain on solid-phase and subsequently converted to ADPr-sites by chemical phosphorylation with adenosine phosphoramidite. Suitably protected phosphoribosylated glutamine, asparagine, and citrulline building blocks described in this protocol allow introduction of ADP-Gln, ADPr-Asn, and ADPr-Cit into peptide chains as demonstrated for three peptides. Trifunctional amino acids, for which base-sensitive side-chain protection is available, can be accommodated in the sequences flanking the ADPr-cites.

Key words

Mono-ADPr-peptides Solid-phase synthesis Pyrophosphate Glycosylation Phosphitylation Phosphoribosylated amino acids 





Acetic acid










4-(N-[1-(4,4-dimethyl-2,6-dioxocyclohexylidene)-3-methylbutyl]amino) benzyl ester




Diethyl ether




O-(1H-6-Chlorobenzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate








tert-butyldimethylsilyl triflate




Trifluroacetamide (protective group)


Trifluoroacetic acid



This work was supported by the Netherlands Organization for Scientific Research (NWO).


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Hans A. V. Kistemaker
    • 1
  • Jim Voorneveld
    • 1
  • Dmitri V. Filippov
    • 1
  1. 1.Gorlaeus LaboratoriesLeiden Institute of Chemistry, Universiteit LeidenLeidenThe Netherlands

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