Cited by 4,683Advanced Proteomics Techniques and ApplicationsMass Spectrometry Techniques and ApplicationsRedox biology and oxidative stress

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Quantitative analysis of complex protein mixtures using isotope-coded affinity tags

Gygi, Steven P., Rist, Beate, Gerber, Scott A., Turecek, Frantisek, Gelb, Michael H., Aebersold, Ruedi·1999·Nature Biotechnology

DOI 10.1038/13690

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Abstract

We describe an approach for the accurate quantification and concurrent sequence identification of the individual proteins within complex mixtures. The method is based on a class of new chemical reagents termed isotope-coded affinity tags (ICATs) and tandem mass spectrometry. Using this strategy, we compared protein expression in the yeast Saccharomyces cerevisiae, using either ethanol or galactose as a carbon source. The measured differences in protein expression correlated with known yeast metabolic function under glucose-repressed conditions. The method is redundant if multiple cysteinyl residues are present, and the relative quantification is highly accurate because it is based on stable isotope dilution techniques. The ICAT approach should provide a widely applicable means to compare quantitatively global protein expression in cells and tissues.

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“(2) Then, all free amines (α- and ε-amines) are acetylated (indicated by filled diamonds) and the proteins are digested with trypsin (3), which will now only cleave C-terminal to arginine residues.”
Background
Shotgun identification of protein modifications from protein complexes and lens tissue
MacCoss, M. J., McDonald, W. H., Saraf, A., Sadygov, R. G., Clark, J., Tasto, J. J., Gould, K. L., Wolters, D. A., Washburn, M. P., Weiss, A., et al. (2002) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.122231399 · Cited by 593
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Mechanism-Based Profiling of Enzyme Families
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Self-citation
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Method
Relative and Absolute Quantification of Postsynaptic Density Proteome Isolated from Rat Forebrain and Cerebellum
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Method
A novel strategy for quantitative proteomics using isotope‐coded protein labels
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Method
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Background
Biomarker Discovery from Pancreatic Cancer Secretome Using a Differential Proteomic Approach
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Background
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Background
Quantitative Atlas of Membrane Transporter Proteins: Development and Application of a Highly Sensitive Simultaneous LC/MS/MS Method Combined with Novel In-silico Peptide Selection Criteria
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Authors: Gygi, S. P., Rist, B., Gerber, S. A., Turecek, F., Gelb, M. H., & Aebersold, R.
Affiliations: University of Washington (US)
Corresponding author: Aebersold, Ruedi
ORCID: Gygi, Steven P.
Gerber, Scott A.
Gelb, Michael H.
Aebersold, Ruedi
Venue: Nature Biotechnology
Publisher: Springer Science and Business Media LLC
Published: 1999-10
Volume: 17
Issue: 10
Pages: 994-999
ISSN: 0733-222X (ISSN-L)
1087-0156
1546-1696
DOI: 10.1038/13690
OpenAlex: W2111208057
Semantic Scholar: 8929f01bc9a788127bcff7a09432f4f7979c8e6b
Cited by: 4,683
References: 24
Citations by year: 2026: 19 · 2025: 27 · 2024: 31 · 2023: 54 · 2022: 50 · 2021: 63 · 2020: 51 · 2019: 70 · 2018: 77 · 2017: 104 · 2016: 123 · 2015: 171 · 2014: 188 · 2013: 235 · 2012: 296
FWCI: 75.18
Type: journal article
Language: en
Open access: closed
Licence: www.springer.com/tdm (tdm)
Locations: Nature Biotechnology (publishedVersion)
PubMed (publishedVersion)
Funders: National Human Genome Research Institute (T32 HG-00035, T32HG000035)
National Center for Research Resources (RR-11823, P41RR011823)
Eunice Kennedy Shriver National Institute of Child Health and Human Development (HD 02274, P30HD002274)
Topics: Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
Keywords: ICAT, protein quantification, tandem mass spectrometry, protein expression profiling, Saccharomyces cerevisiae, glucose repression, ethanol carbon source, stable isotope dilution, cysteine labeling
MeSH terms: Affinity Labels, Amino Acid Sequence, Chromatography, Liquid, Isotope Labeling, Proteins, Mass Spectrometry

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Quantitative analysis of complex protein mixtures using isotope-coded affinity tags

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