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Peptide Dimethylation: Fragmentation Control via Distancing the Dimethylamino Group

  • Research Article
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Journal of The American Society for Mass Spectrometry

Abstract

Direct reductive methylation of peptides is a common method for quantitative proteomics. It is an active derivatization technique; with participation of the dimethylamino group, the derivatized peptides preferentially release intense a1 ions. The advantageous generation of a1 ions for quantitative proteomic profiling, however, is not desirable for targeted proteomic quantitation using multiple reaction monitoring mass spectrometry; this mass spectrometric method prefers the derivatizing group to stay with the intact peptide ions and multiple fragments as passive mass tags. This work investigated collisional fragmentation of peptides whose amine groups were derivatized with five linear ω-dimethylamino acids, from 2-(dimethylamino)-acetic acid to 6-(dimethylamino)-hexanoic acid. Tandem mass spectra of the derivatized tryptic peptides revealed different preferential breakdown pathways. Together with energy resolved mass spectrometry, it was found that shutting down the active participation of the terminal dimethylamino group in fragmentation of derivatized peptides is possible. However, it took a separation of five methylene groups between the terminal dimethylamino group and the amide formed upon peptide derivatization. For the first time, the gas-phase fragmentation of peptides derivatized with linear ω-dimethylamino acids of systematically increasing alkyl chain lengths is reported.

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Acknowledgments

The authors greatly appreciate financial support from the Cystic Fibrosis Foundation (YAO07XX0) and the NCI/NIH (1R21CA155536-01). They thank Song Li for his assistance with NMR spectroscopy and the Leadbeater Lab for use of their scientific microwave.

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Authors and Affiliations

  1. Department of Chemistry, University of Connecticut, Storrs, CT, 06269, USA

    Adam J. McShane, Yuanyuan Shen, Mary Joan Castillo & Xudong Yao

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  1. Adam J. McShane
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  2. Yuanyuan Shen
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Correspondence to Xudong Yao.

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McShane, A.J., Shen, Y., Castillo, M.J. et al. Peptide Dimethylation: Fragmentation Control via Distancing the Dimethylamino Group. J. Am. Soc. Mass Spectrom. 25, 1694–1704 (2014). https://doi.org/10.1007/s13361-014-0951-7

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  • DOI: https://doi.org/10.1007/s13361-014-0951-7

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