Scientific article

N-t-butyliodoacetamide and iodoacetanilide: two new cysteine alkylating reagents for relative quantitation of proteins

Published inRCM. Rapid communications in mass spectrometry, vol. 18, no. 1, p. 117-127
Publication date2004

The synthesis and application of two new alkylating reagents, N-tert-butyl-2-iodoacetamide (N-t-butyliodoacetamide) and 2-iodo-N-phenylacetamide (iodoacetanilide), are described. N-t-Butyliodoacetamide and iodoacetanilide were synthesised to purity in their d(0)-light and in their respective d(9)- and d(5)-heavy forms. The newly synthesised reagents are covalently bound to peptides containing cysteines via an alkylation reaction. The mass differences of 5 and 9 Da avoid possible problems of overlapping isotope distribution. For each alkylated cysteine a peptide mass increases, respectively, by a multiple of 113 and 133 Da for the d(0)-light form of N-t-butyliodoacetamide and iodoacetanilide. These reagents can therefore replace common alkylating reagents in existing proteomics-based applications. Alkylated peptides increase in mass in the same mass range as amino acids and remain suitable for tandem mass spectrometry (MS/MS) data acquisition and analysis. The compounds are simple to use and derivatisation is based on widely applied alkylating procedures. Preliminary results show that these reagents can be applied for both protein quantitation and identification by peptide mass finger printing and/or MS/MS techniques. Using these chemicals and the suggested workflow enables the quantitative analysis of the whole protein sample and realises access to peptides that may contain potential post-translational modifications. Other approaches that incorporate a matrix-assisted laser desorption/ionisation (MALDI) interface prior to MS can take advantage of these chemicals, such as the molecular scanner.

  • Acetanilides/chemistry
  • Alkylating Agents/chemistry
  • Alkylation
  • Cysteine/chemistry
  • Indicators and Reagents/chemistry
  • Iodoacetamide/chemistry
  • Lactoperoxidase/analysis/chemistry
  • Peptide Mapping/methods
  • Proteins/analysis/chemistry
  • Proteomics/methods
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
Citation (ISO format)
PASQUARELLO MOSIMANN, Carla et al. N-t-butyliodoacetamide and iodoacetanilide: two new cysteine alkylating reagents for relative quantitation of proteins. In: RCM. Rapid communications in mass spectrometry, 2004, vol. 18, n° 1, p. 117–127. doi: 10.1002/rcm.1286
Main files (1)
Article (Published version)
ISSN of the journal0951-4198

Technical informations

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