Development of activity-based probes for trypsin-family serine proteases

• Published in: Bioorganic & medicinal chemistry letters Vol. 16; no. 11; pp. 2882 - 2885
• Main Authors: Pan, Zhengying, Jeffery, Douglas A., Chehade, Kareem, Beltman, Jerlyn, Clark, James M., Grothaus, Paul, Bogyo, Matthew, Baruch, Amos
• Format: Journal Article
• Language: English
• Published: Oxford Elsevier Ltd 01.06.2006; Elsevier
• Subjects: Activity-based probes; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Cell Line; Chemical proteomics; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; Humans; Mast Cells - drug effects; Mast Cells - enzymology; Molecular Structure; Proteins; Trypsin - classification; Trypsin - metabolism; Trypsin Inhibitors - chemical synthesis; Trypsin Inhibitors - chemistry; Trypsin Inhibitors - pharmacology; Tryptase; Activity-based probes; Chemical proteomics; Tryptase; Biotinylation; Serine endopeptidases; Peptides; Enzyme; Gel electrophoresis; Organic phosphonate; Selectivity; Chemical labelling; Biological activity; Phosphorus Organic compounds; Ethylene oxide polymer; Peptidases; Trypsin; Enzymatic activity; Proteomics; Dipeptides; Hydrolases; Mast cell; Detection
• ISSN: 0960-894X; 1464-3405
• DOI: 10.1016/j.bmcl.2006.03.012

Development and applications of a series of diphenylphosphonate-based probes for the trypsin-like serine proteases are reported. A series of diphenylphosphonate-based probes were developed for the trypsin-like serine proteases. These probes selectively target serine proteases rather than general serine hydrolases that are targets for fluorophosphonate-based probes. This increased selectivity allows detection of low abundance serine proteases in complex proteomes using simple SDS–PAGE methods. We present here the application of multiple probes in enzyme activity profiling of intact mast cells, a type of inflammatory cell implicated in allergy and autoimmune diseases.