Quantitative peptidomics using reductive methylation of amines

Sayani Dasgupta, Leandro M. Castro, Alexandre K. Tashima, Lloyd D. Fricker

Research output: Chapter in Book/Report/Conference proceedingChapter

1 Scopus citations

Abstract

A number of different approaches have been used for quantitative peptidomics. In this protocol we describe the method in which peptides are reacted with formaldehyde and sodium cyanoborohydride, which converts primary and secondary amines into tertiary amines. By using different combinations of regular reagents, deuterated reagents (2H), and reagents containing deuterium and 13C, it is possible to produce five isotopically distinct forms of the methylated peptides which can be quantified by mass spectrometry. Peptides with free N-termini that are primary amines incorporate two methyl groups using this procedure, which differ by 2 Da for each of the five isotopic combinations. Peptides that contain unmodified lysine residues incorporate additional pairs of methyl groups, leading to larger mass differences between isotopic forms. The reagents are commercially available, relatively inexpensive, and chemically stable.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages161-174
Number of pages14
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1719
ISSN (Print)1064-3745

Keywords

  • Cyanoborohydride
  • Formaldehyde
  • Isotopic labels

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Dasgupta, S., Castro, L. M., Tashima, A. K., & Fricker, L. D. (2018). Quantitative peptidomics using reductive methylation of amines. In Methods in Molecular Biology (pp. 161-174). (Methods in Molecular Biology; Vol. 1719). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7537-2_10