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Fiona J Flett, Timo Sachsenberg, Oliver Kohlbacher, C. L Mackay, and Heidrun Interthal (in press)

Differential Enzymatic 16O/18O Labelling for the Detection of Cross-Linked Nucleic Acid-Protein Heteroconjugates

Anal. Chem., in press.

Cross-linking of nucleic acids to proteins in combination with mass spectrometry permits the precise identification of interacting residues between nucleic acid-protein complexes. However, the mass spectrometric identification and characterisation of cross-linked nucleic acid-protein heteroconjugates within a complex sample is challenging. Here we establish a novel enzymatic differential 16O/18O labelling approach, which uniquely labels heteroconjugates. We have developed an automated data analysis workflow based on OpenMS for the identification of differentially isotopically labelled heteroconjugates against a complex background. We validated our method using synthetic model DNA oligonucleotide-peptide heteroconjugates which were subjected to the labelling reaction and analysed by high resolution FT-ICR mass spectrometry.
Sachsenberg, Timo; Kohlbacher, Oliver; Mackay, C. Logan; Interthal, Heidrun
28885003
10.1021/acs.analchem.7b01625