title: Phosphoramidates as novel activity-based probes for serine proteases.
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.
authors.affiliation: Lehrstuhl für Chemie der Biopolymere, Technische Universität München, Weihenstephaner Berg 3, 85354 Freising (Germany).
authors.forename: Ute R
authors.initials: UR
authors.lastname: Haedke
authors.forename: Sandra C
authors.initials: SC
authors.lastname: Frommel
authors.forename: Fabian
authors.initials: F
authors.lastname: Hansen
authors.forename: Hannes
authors.initials: H
authors.lastname: Hahne
authors.forename: Bernhard
authors.initials: B
authors.lastname: Kuster
authors.forename: Matthew
authors.initials: M
authors.lastname: Bogyo
authors.forename: Steven H L
authors.initials: SH
authors.lastname: Verhelst
mesh_terms: D000577:Amides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D008968:Molecular Conformation; D015336:Molecular Probe Techniques; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D010756:Phosphoric Acids / Q000138:chemical synthesis / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D057057:Serine Proteases / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000577:Amides; D015335:Molecular Probes; D010455:Peptides; D010756:Phosphoric Acids; C011067:phosphoramidic acid; D057057:Serine Proteases
keywords: activity-based probes; proteases; proteomics; solid-phase synthesis; target identification