Click Chemistry Generated Model DNA鈥揚eptide Heteroconjugates as Tools for Mass Spectrometry

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• 作者：Fiona J. Flett ; Jeffrey G. A. Walton ; C. Logan Mackay ; Heidrun Interthal
• 刊名：Analytical Chemistry
• 出版年：2015
• 出版时间：October 6, 2015
• 年：2015
• 卷：87
• 期：19
• 页码：9595-9599
• 全文大小：303K
• ISSN：1520-6882

文摘

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鈥損rotein complexes. However, the mass spectrometric identification of cross-linked nucleic acid鈥損rotein 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鈥損eptide 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鈥損eptide heteroconjugates have the potential to be of use for a variety of applications that involve DNA鈥損eptide heteroconjugates.