Chemoproteomic profiling and discovery of protein electrophiles in human cells
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Activity-based protein profiling (ABPP) serves as a chemical proteomic platform to discover and characterize functional amino acids in proteins on the basis of their enhanced reactivity towards small-molecule probes. This approach, to date, has mainly targeted nucleophilic functional groups, such as the side chains of serine and cysteine, using electrophilic probes. Here we show that 'reverse-polarity' (RP)-ABPP using clickable, nucleophilic hydrazine probes can capture and identify protein-bound electrophiles in cells. Using this approach, we demonstrate that the pyruvoyl cofactor of S-adenosyl-L-methionine decarboxylase (AMD1) is dynamically controlled by intracellular methionine concentrations. We also identify a heretofore unknown modification-an N-terminally bound glyoxylyl group-in the poorly characterized protein secernin-3. RP-ABPP thus provides a versatile method to monitor the metabolic regulation of electrophilic cofactors and discover novel types of electrophilic modifications on proteins in human cells.
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ABPP principles and reverse-polarity probes have been employed to study electrophilic post-translational modifications such as N-terminal pyruvoyl and glyoxylyl modifications that can react with hydrazine-containing compounds.
“ABPP principles and ‘reverse-polarity’ probes have been employed to study electrophilic post-translational modifications, such as N-terminal pyruvoyl and glyoxylyl modifications, that can react with hydrazine-containing compounds197.”
From Advances in covalent drug discovery (2022) · cites this paper as [197]
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- Authors
- Matthews, M. L., He, L., Horning, B. D., Olson, E. J., Correia, B. E., Yates, J. R., Dawson, P. E., & Cravatt, B. F.
- Affiliations
- Scripps Research Institute (US)
- Corresponding author
- Matthews, Megan L.
- Cravatt, Benjamin F.
- ORCID
- Venue
- Nature Chemistry
- Published
- 2016-10-31
- Volume
- 9
- Issue
- 3
- Pages
- 234-243
- ISSN
- 1755-4330 (ISSN-L)
- 1755-4349
- PMC
- PMC5325178
- OpenAlex
- W2542506435
- Cited by
- 90
- References
- 54
- Citations by year
- 2026: 4 · 2025: 11 · 2024: 8 · 2023: 10 · 2022: 11 · 2021: 11 · 2020: 12 · 2019: 8 · 2018: 8 · 2017: 6 · 2016: 1
- FWCI
- 3.52
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Chemistry (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Infoscience (Ecole Polytechnique Fédérale de Lausanne) (submittedVersion)
- Infoscience (Ecole Polytechnique Fédérale de Lausanne) (submittedVersion, cc-by-nc-nd)
- Funders
- National Cancer Institute (R01CA132630)
- National Institute of General Medical Sciences (P41GM103533, U54GM114833)
- Topics
- Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Keywords
- activity-based protein profiling, S-adenosylmethionine decarboxylase, methionine metabolism, protein modification, cofactor regulation, human cells
- MeSH terms
- Amino Acids, Cell Survival, Fluorescent Dyes, Humans, Hydrazines, Methionine, Nerve Tissue Proteins, Adenosylmethionine Decarboxylase, Molecular Structure, Proteomics, HEK293 Cells
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