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Median-Based Absolute Quantification of Proteins Using Fully Unlabeled Generic Internal Standard (FUGIS)
Open accessPublished version of a preprint
Abstract
By reporting the molar abundance of proteins, absolute quantification determines their stoichiometry in complexes, pathways, or networks. Typically, absolute quantification relies either on protein-specific isotopically labeled peptide standards or on a semiempirical calibration against the average abundance of peptides chosen from arbitrarily selected proteins. In contrast, a generic protein standard FUGIS (fully unlabeled generic internal standard) requires no isotopic labeling, chemical synthesis, or external calibration and is applicable to quantifying proteins of any organismal origin. The median intensity of the peptide peaks produced by the tryptic digestion of FUGIS is used as a single-point calibrant to determine the molar abundance of any codigested protein. Powered by FUGIS, median-based absolute quantification (MBAQ) outperformed other methods of untargeted proteome-wide absolute quantification.
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Cites
Global quantification of mammalian gene expression control
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A “Proteomic Ruler” for Protein Copy Number and Concentration Estimation without Spike-in Standards
Wiśniewski, J. R., Hein, M. Y., Cox, J., & Mann, M. (2014) · Molecular & Cellular Proteomics · doi:10.1074/mcp.m113.037309 · Cited by 690
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Metadata
- Authors
- Raghuraman, B. K., Bogdanova, A., Moon, H., Rzagalinski, I., Geertsma, E. R., Hersemann, L., & Shevchenko, A.
- Affiliations
- Max Planck Institute of Molecular Cell Biology and Genetics (DE)
- Corresponding author
- Shevchenko, Andrej
- Venue
- Journal of Proteome Research
- Publisher
- American Chemical Society (ACS)
- Published
- 2021-11-22
- Volume
- 21
- Issue
- 1
- Pages
- 132-141
- ISSN
- 1535-3893 (ISSN-L)
- 1535-3907
- PMC
- PMC8749952
- OpenAlex
- W3217139745
- Cited by
- 9
- References
- 3
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 2 · 2022: 1 · 2021: 1
- FWCI
- 0.37
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- Journal of Proteome Research (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Max-Planck-Gesellschaft
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
- Keywords
- absolute protein quantification, trypsin digestion
- MeSH terms
- Calibration, Isotope Labeling, Peptides, Reference Standards, Proteome
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