Application of a Sulfoxonium Ylide Electrophile to Generate Cathepsin X-Selective Activity-Based Probes
| Autor: | Simon J. Mountford, Jiayin Diao, Bangyan Xu, Bethany M. Anderson, Erik Lindström, Philip E. Thompson, Elean S.V. Tay, Luigi Aurelio, Erica K. Sloan, Robin M. Yates, Monika Szabo, Rhiannon I. Campden, Nigel W. Bunnett, Laura E. Edgington-Mitchell, My Linh Hoang |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Male Hydrocarbons Fluorinated Cell Culture Techniques Kidney 01 natural sciences Biochemistry Article Cell Line Substrate Specificity Carboxypeptidase activity Small Molecule Libraries 03 medical and health sciences Mice Structure-Activity Relationship Protein Domains In vivo Structure–activity relationship Animals Humans Amino Acids Enzyme Inhibitors Fluorescent Dyes Cathepsin chemistry.chemical_classification 010405 organic chemistry Chemistry Optical Imaging General Medicine Ketones Cathepsins 0104 chemical sciences 3. Good health Amino acid Mice Inbred C57BL Kinetics 030104 developmental biology Diazomethane Cell culture Molecular Medicine Function (biology) Cysteine |
| Zdroj: | ACS Chem Biol |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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