Click chemistry generated model DNA-Peptide Heteroconjugates as tools for mass spectrometry

Fiona J. Flett, Jeffrey G A Walton, C. Logan Mackay, Heidrun Interthal*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

UV cross-linking of nucleic acids to proteins in combination with mass spectrometry is a powerful technique to identify proteins, peptides, and the amino acids involved in intermolecular interactions within nucleic acid-protein complexes. However, the mass spectrometric identification of cross-linked nucleic acid-protein heteroconjugates in complex mixtures and MS/MS characterization of the specific sites of cross-linking is extremely challenging. As a tool for the optimization of sample preparation, ionization, fragmentation, and detection by mass spectrometry, novel synthetic DNA-peptide heteroconjugates were generated to act as mimics of UV cross-linked heteroconjugates. Click chemistry was employed to cross-link peptides to DNA oligonucleotides. These heteroconjugates were fully characterized by high resolution FTICR mass spectrometry and by collision-induced dissociation (CID) following nuclease P1 digestion of the DNA moiety to a single nucleotide monophosphate. This allowed the exact site of the cross-linking within the peptide to be unambiguously assigned. These synthetic DNA-peptide heteroconjugates have the potential to be of use for a variety of applications that involve DNA-peptide heteroconjugates.

Original languageEnglish
Pages (from-to)9595-9599
Number of pages5
JournalAnalytical Chemistry
Issue number19
Publication statusPublished - 3 Sept 2015


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