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Phosphoramidates as Novel Activity‐Based Probes for Serine Proteases
Author(s) -
Haedke Ute R.,
Frommel Sandra C.,
Hansen Fabian,
Hahne Hannes,
Kuster Bernhard,
Bogyo Matthew,
Verhelst Steven H. L.
Publication year - 2014
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201400013
Subject(s) - proteases , serine , proteome , biochemistry , chemistry , enzyme , proteomics , biology , gene
Activity‐based probes (ABPs) are small molecules that exclusively form covalent bonds with catalytically active enzymes. In the last decade, they have especially been used in functional proteomics studies of proteases. Here, we present phosphoramidate peptides as a novel type of ABP for serine proteases. These molecules can be made in a straightforward manner by standard Fmoc‐based solid‐phase peptide synthesis, allowing rapid diversification. The resulting ABPs covalently bind different serine proteases, depending on the amino acid recognition element adjacent to the reactive group. A reporter tag enables downstream gel‐based analysis or LC‐MS/MS‐mediated identification of the targeted proteases. Overall, we believe that these readily accessible probes will provide new avenues for the functional study of serine proteases in complex proteomes.