Synthesis of Mono-ADP-Ribosylated Oligopeptides Using Ribosylated Amino Acid Building Blocks

Abstract

Adenosine diphosphate ribosylation (ADP-ribosylation) is a widely occurring post-translational modification of proteins at nucleophilic side chains of amino acid residues, such as asparagine, glutamic acid, and arginine. Elucidation of the biological role of ADP-ribosylation events would benefit from the availability of well-defined ADP-ribosylated peptides. Main issues in the construction of synthetic ADP-ribosylated peptides involve the availability of protected ribosylated amino acids suitable for peptide synthesis, development of a protective group strategy for peptide fragments compatible with the integrity of the adenosine diphosphate moiety, and an efficient procedure for pyrophosphate formation. In this paper we present a first approach to the chemical synthesis of ADP-ribosylated peptides in solution and on solid support. We describe an efficient synthesis of suitably protected ribosylated asparagine and glutamine building blocks suitable for Fmoc-based peptide synthesis. We further demonstrate a successful application of these ribosylated amino acids in the assembly of three fully synthetic ADP-ribosylated peptides by solution and solid phase approaches.

Chemical Methods for Encoding and Decoding of Posttranslational Modifications

“…However, the chemically sensitive nature of the pyrophosphate bond makes peptide synthesis challenging. In order to circumvent these issues, Filippov and co-workers installed a selectively protected ribosylated asparagine or glutamine residue into peptide using solid phase peptide synthesis (van der Heden van Noort et al, 2010). An alternative approach has also been developed for the generation of ADP-ribosylation analogs (Figure 7F) (Moyle and Muir, 2010).…”

Poly(ADP-Ribose)—A Unique Natural Polymer Structural Features, Biological Role and Approaches to the Chemical Synthesis

“…[59] Modifications of histones by ADPR may be valuable instrument for investigation of protein interactions with PAR and mechanisms of PARP, PARG, and ADP-ribosylproteinlyase action in vitro. Methods of peptide chemistry can be recruited for such modifications.…”

“…As an example, ADP-ribosylated isosterical fragment of histone H2B was obtained by this way (Scheme 6). [59] This fragment is located at the N -terminal part of histone and undergoes PAR-ylation during DNA replication or repair. [59] Glutamine, containing ribose moiety (compound A), was included into heptapeptide by peptide chemistry.…”

“…[59] This fragment is located at the N -terminal part of histone and undergoes PAR-ylation during DNA replication or repair. [59] Glutamine, containing ribose moiety (compound A), was included into heptapeptide by peptide chemistry. After selective removal of tert-butyldiphenylsilyl (TBDPS) protecting group, followed by phosphorylation, the resulting peptide reacted with activated nucleotide B. Deprotection and cleavage from the polymeric carrier was carried out in NH 3 /MeOH.…”

A review of tandem mass spectrometry characterization of adenosine diphosphate-ribosylated peptides

“…Recently, two routes of chemical synthesis have been proposed to make relatively pure ADP-ribosylated peptide, or protein, standards without the use of an enzyme. The first approach involves synthesis of protected ribosylated asparagines and glutamine building blocks that are incorporated into peptides via solid phase synthesis, and then undergo reaction to generate ADP-ribosylated peptides [23]. The second method uses aminooxy functionalized amino acids for the incorporation of ADP-ribose onto specific amino acid functional groups [24].…”