Analytical Chemistry
Analytical Chemistry
4 years ago

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

Differential Enzymatic 16O/18O Labeling for the Detection of Cross-Linked Nucleic Acid–Protein Heteroconjugates
C. Logan Mackay, Fiona J. Flett, Heidrun Interthal, Oliver Kohlbacher, Timo Sachsenberg
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 characterization of cross-linked nucleic acid–protein heteroconjugates within a complex sample is challenging. Here we establish a novel enzymatic differential 16O/18O-labeling approach, which uniquely labels heteroconjugates. We have developed an automated data analysis workflow based on OpenMS for the identification of differentially isotopically labeled heteroconjugates against a complex background. We validated our method using synthetic model DNA oligonucleotide–peptide heteroconjugates, which were subjected to the labeling reaction and analyzed by high-resolution FTICR mass spectrometry.

Publisher URL: http://dx.doi.org/10.1021/acs.analchem.7b01625

DOI: 10.1021/acs.analchem.7b01625

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