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

Flett, Fiona J.; Sachsenberg, Timo; Kohlbacher, Oliver; Mackay, C. Logan; Interthal, Heidrun

doi:10.1021/acs.analchem.7b01625

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

MPS-Authors

Kohlbacher, Oliver
Center for Bioinformatics, Quantitative Biology Center and Department of Computer Science, University of Tübingen;
Max Planck Institute for Developmental Biology, Max Planck Society;

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https://pubs.acs.org/doi/10.1021/acs.analchem.7b01625
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Citation

Flett, F. J., Sachsenberg, T., Kohlbacher, O., Mackay, C. L., & Interthal, H. (2017). Differential Enzymatic O-16/O-18 Labeling for the Detection of Cross-Linked Nucleic Acid-Protein Heteroconjugates. ANALYTICAL CHEMISTRY, 89(21), 11208-11213. doi:10.1021/acs.analchem.7b01625.

Cite as: https://hdl.handle.net/21.11116/0000-0002-1A42-7

Abstract

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 O-16/O-18-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.