Abstract
An important goal in proteomics is to compare the relative amounts of different proteins in biological samples and to try to correlate these differences with changes in physiological state. The isotope- coded affinity tag technique pioneered in Aebersold’s laboratory takes advantage of differential tagging of cysteine residues in proteins with stable isotopes to significantly reduce the complexity of peptide mixtures and increase the number of sequences that are identified in a single tandem mass spectrometry experiment. In this approach, two samples are isotopically labeled (one heavy, one light) through a reactive group that specifically binds to cysteine residues; the samples are combined, separated with chromatography, and analyzed by mass spectrometry. The results are then database searched and a list of hundreds of proteins and their heavy:light ratio is obtained.
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References
Gygi S. P., Rist B., Gerber S. A., Turecek F., Gelb M. H., and Aebersold R. (1999) Quantitative analysis of complex protein mixtures using isotope coded affinity tags. Nat. Biotechnol. 17, 994–999.
Han D. K., Eng J., Zhou H., and Aebersold R. (2001) Quantitative profiling of differentiation-induced microsomal proteins using isotope-coded affinity tags and mass spectrometry. Nat Biotechnol. 19, 946–951.
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Colangelo, C.M., Williams, K.R. (2006). Isotope-Coded Affinity Tags for Protein Quantification. In: New and Emerging Proteomic Techniques. Methods in Molecular Biology™, vol 328. Humana Press. https://doi.org/10.1385/1-59745-026-X:151
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DOI: https://doi.org/10.1385/1-59745-026-X:151
Publisher Name: Humana Press
Print ISBN: 978-1-58829-519-4
Online ISBN: 978-1-59745-026-3
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