Title | A Chemical Strategy for Protease Substrate Profiling. |
Publication Type | Journal Article |
Year of Publication | 2019 |
Authors | Griswold AR, Cifani P, Rao SD, Axelrod AJ, Miele MM, Hendrickson RC, Kentsis A, Bachovchin DA |
Journal | Cell Chem Biol |
Volume | 26 |
Issue | 6 |
Pagination | 901-907.e6 |
Date Published | 2019 Jun 20 |
ISSN | 2451-9448 |
Abstract | The dipeptidyl peptidases (DPPs) regulate hormones, cytokines, and neuropeptides by cleaving dipeptides after proline from their amino termini. Due to technical challenges, many DPP substrates remain unknown. Here, we introduce a simple method, termed CHOPS (chemical enrichment of protease substrates), for the discovery of protease substrates. CHOPS exploits a 2-pyridinecarboxaldehyde (2PCA)-biotin probe, which selectively biotinylates protein N-termini except those with proline in the second position. CHOPS can, in theory, discover substrates for any protease, but is particularly well suited to discover canonical DPP substrates, as cleaved but not intact DPP substrates can be identified by gel electrophoresis or mass spectrometry. Using CHOPS, we show that DPP8 and DPP9, enzymes that control the Nlrp1 inflammasome through an unknown mechanism, do not directly cleave Nlrp1. We further show that DPP9 robustly cleaves short peptides but not full-length proteins. More generally, this work delineates a practical technology for identifying protease substrates, which we anticipate will complement available "N-terminomic" approaches. |
DOI | 10.1016/j.chembiol.2019.03.007 |
Alternate Journal | Cell Chem Biol |
PubMed ID | 31006619 |
PubMed Central ID | PMC6588500 |
Grant List | R01 CA204396 / CA / NCI NIH HHS / United States R01 AI137168 / AI / NIAID NIH HHS / United States P30 CA008748 / CA / NCI NIH HHS / United States R21 CA188881 / CA / NCI NIH HHS / United States / / Wellcome Trust / United Kingdom T32 GM007739 / GM / NIGMS NIH HHS / United States |
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