Quantitative Enzyme Activity Analysis by ABPP and LC-MS/MS
- Only active enzymes react with warheads.
- Include competition controls.
- Probe class defines enzyme coverage.
- MS quant tracks activity changes under drug treatment.
Incubate lysates with ABPs, enrich tagged enzymes, digest, and quantify probe-modified peptides by label-free or TMT LC-MS/MS across conditions.
Key Takeaways
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FAQ
1. ABPP vs expression proteomics?
ABPP reports catalytic activity, not total protein level.
2. What are limitations?
Probe design and non-covalent enzymes need tailored strategies.
Conclusion
Quantitative ABPP connects functional enzyme states to phenotypes and compound action.
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