1. Application of a Sulfoxonium Ylide Electrophile to Generate Cathepsin X-Selective Activity-Based Probes.
- Author
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Mountford SJ, Anderson BM, Xu B, Tay ESV, Szabo M, Hoang ML, Diao J, Aurelio L, Campden RI, Lindström E, Sloan EK, Yates RM, Bunnett NW, Thompson PE, and Edgington-Mitchell LE
- Subjects
- Amino Acids chemistry, Animals, Cell Culture Techniques, Cell Line, Diazomethane chemistry, Humans, Hydrocarbons, Fluorinated chemistry, Ketones chemistry, Kidney cytology, Kidney diagnostic imaging, Kinetics, Male, Mice, Mice, Inbred C57BL, Optical Imaging, Protein Domains, Small Molecule Libraries chemistry, Structure-Activity Relationship, Substrate Specificity, Cathepsins antagonists & inhibitors, Enzyme Inhibitors chemical synthesis, Enzyme Inhibitors metabolism, Fluorescent Dyes chemistry
- Abstract
Cathepsin X/Z/P is cysteine cathepsin with unique carboxypeptidase activity. Its expression is associated with cancer and neurodegenerative diseases, although its roles during normal physiology are still poorly understood. Advances in our understanding of its function have been hindered by a lack of available tools that can specifically measure the proteolytic activity of cathepsin X. We present a series of activity-based probes that incorporate a sulfoxonium ylide warhead, which exhibit improved specificity for cathepsin X compared to previously reported probes. We apply these probes to detect cathepsin X activity in cell and tissue lysates, in live cells and in vivo, and to localize active cathepsin X in mouse tissues by microscopy. Finally, we utilize an improved method to generate chloromethylketones, necessary intermediates for synthesis of acyloxymethylketones probes, by way of sulfoxonium ylide intermediates. In conclusion, the probes presented in this study will be valuable for investigating cathepsin X pathophysiology.
- Published
- 2020
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