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

作者：Flett Fiona J; Sachsenberg Timo; Kohlbacher Oliver; Mackay C Logan; Interthal Heidrun^*

来源：Analytical Chemistry, 2017, 89(21): 11208-11213.

DOI：10.1021/acs.analchem.7b01625

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.

• 出版日期2017-11-7
• 单位上海生物信息技术研究中心