Phosphoramidates as Novel Activity‐Based Probes for Serine Proteases |
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| Authors: | Dr. Ute R. Haedke Sandra C. Frommel Fabian Hansen Dr. Hannes Hahne Prof. Dr. Bernhard Kuster Prof. Dr. Matthew Bogyo Dr. Steven H. L. Verhelst |
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| Affiliation: | 1. Lehrstuhl für Chemie der Biopolymere, Technische Universit?t München, Weihenstephaner Berg 3, 85354 Freising (Germany);2. Chair of Proteomics and Bioanalytics, Technische Universit?t München, Emil Erlenmeyer Forum 5, 85354 Freising (Germany);3. Departments of Pathology and Microbiology & Immunology, Stanford University, School of Medicine, 300 Pasteur Drive, CA 94305 Stanford (USA) |
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| Abstract: | 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. |
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| Keywords: | activity‐based probes proteases proteomics solid‐phase synthesis target identification |
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