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Selectivity of labeled bromoethylamine for protein alkylation

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Abstract

Alkylation of cysteine residues has been used extensively for characterization of proteins and their mode of action in biological systems, research endeavors that are at the core of proteomics. Treatment with a simple alkylating agent such as [2-13C] bromoethylamine would result in labeled thialysine at the ε-position. This chemical modification of proteins would allow investigations via both 13C NMR spectroscopy and mass spectrometry. However [2-13C] labeled bromoethylamine is not available commercially. We investigated its synthesis at acid pH with the goal of obtaining singly labeled bromoethylamine and understanding the mechanistic details of the reaction. Based on our experimental and theoretical results, bromination of [2-13C] labeled ethanolamine in acidic conditions takes place via exclusive attack of the nucleophile (HBr) at the hydroxyl bearing C. Moreover, hydrogen bonding guides the nucleophilic attack, resulting in no label scrambling of the bromoethylamine product. Protein alkylation at cysteine residue with the synthesized Br13CH2CH2NH2-HBr is successful. Ab initio calculations in which CH3SH serves as a model for the cysteine residue suggest that in gas phase intermolecular attack by the sulfur bearing nucleophile is favored over the intramolecular substitution by the amino group by 15.4 kJ mol−1. Solution modeling shows that the trend is preserved at basic pH, which is the experimental one, but is reversed at neutral pH.

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Authors and Affiliations

  1. Department of Natural Sciences, University of Michigan-Dearborn, 4901 Evergreen Rd., Dearborn, MI, 48128, USA

    Simona Marincean

  2. Department of Chemistry and Biochemistry, Kettering University, 1700 University Ave, Flint, MI, 48504, USA

    Montserrat Rabago Smith & Laci Beltz

  3. Department of Chemistry, Michigan State University, East Lansing, MI, 48824, USA

    Babak Borhan

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  1. Simona Marincean
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  2. Montserrat Rabago Smith
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  3. Laci Beltz
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  4. Babak Borhan
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Correspondence to Simona Marincean.

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Marincean, S., Rabago Smith, M., Beltz, L. et al. Selectivity of labeled bromoethylamine for protein alkylation. J Mol Model 18, 4547–4556 (2012). https://doi.org/10.1007/s00894-012-1461-9

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  • DOI: https://doi.org/10.1007/s00894-012-1461-9

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