Detection of ubiquitin-proteasome enzymatic activities in cells: application of activity-based probes to inhibitor development.
BACKGROUND: Synthetic probes that mimic natural substrates can enable the detection of enzymatic activities in a cellular environment. One area where such activity-based probes have been applied is the ubiquitin-proteasome pathway, which is emerging as an important therapeutic target. A family of re...
| Autores principales: | , , , |
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| Formato: | Journal article |
| Lenguaje: | English |
| Publicado: |
2012
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| _version_ | 1826264240779952128 |
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| author | Kramer, H Nicholson, B Kessler, B Altun, M |
| author_facet | Kramer, H Nicholson, B Kessler, B Altun, M |
| author_sort | Kramer, H |
| collection | OXFORD |
| description | BACKGROUND: Synthetic probes that mimic natural substrates can enable the detection of enzymatic activities in a cellular environment. One area where such activity-based probes have been applied is the ubiquitin-proteasome pathway, which is emerging as an important therapeutic target. A family of reagents has been developed that specifically label deubiquitylating enzymes (DUBs) and facilitate characterization of their inhibitors. SCOPE OF REVIEW: Here we focus on the application of probes for intracellular DUBs, a group of specific proteases involved in the ubiquitin proteasome system. In particular, the functional characterization of the active subunits of this family of proteases that specifically recognize ubiquitin and ubiquitin-like proteins will be discussed. In addition we present the potential and design of activity-based probes targeting kinases and phosphatases to study phosphorylation. MAJOR CONCLUSIONS: Synthetic molecular probes have increased our understanding of the functional role of DUBs in living cells. In addition to the detection of enzymatic activities of known members, activity-based probes have contributed to a number of functional assignments of previously uncharacterized enzymes. This method enables cellular validation of the specificity of small molecule DUB inhibitors. GENERAL SIGNIFICANCE: Molecular probes combined with mass spectrometry-based proteomics and cellular assays represent a powerful approach for discovery and functional validation, a concept that can be expanded to other enzyme classes. This addresses a need for more informative cell-based assays that are required to accelerate the drug development process. This article is part of a Special Issue entitled: Ubiquitin Drug Discovery and Diagnostics. |
| first_indexed | 2024-03-06T20:04:36Z |
| format | Journal article |
| id | oxford-uuid:28821d2e-24eb-4ce1-998b-3ee951c7132f |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T20:04:36Z |
| publishDate | 2012 |
| record_format | dspace |
| spelling | oxford-uuid:28821d2e-24eb-4ce1-998b-3ee951c7132f2022-03-26T12:13:11ZDetection of ubiquitin-proteasome enzymatic activities in cells: application of activity-based probes to inhibitor development.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:28821d2e-24eb-4ce1-998b-3ee951c7132fEnglishSymplectic Elements at Oxford2012Kramer, HNicholson, BKessler, BAltun, MBACKGROUND: Synthetic probes that mimic natural substrates can enable the detection of enzymatic activities in a cellular environment. One area where such activity-based probes have been applied is the ubiquitin-proteasome pathway, which is emerging as an important therapeutic target. A family of reagents has been developed that specifically label deubiquitylating enzymes (DUBs) and facilitate characterization of their inhibitors. SCOPE OF REVIEW: Here we focus on the application of probes for intracellular DUBs, a group of specific proteases involved in the ubiquitin proteasome system. In particular, the functional characterization of the active subunits of this family of proteases that specifically recognize ubiquitin and ubiquitin-like proteins will be discussed. In addition we present the potential and design of activity-based probes targeting kinases and phosphatases to study phosphorylation. MAJOR CONCLUSIONS: Synthetic molecular probes have increased our understanding of the functional role of DUBs in living cells. In addition to the detection of enzymatic activities of known members, activity-based probes have contributed to a number of functional assignments of previously uncharacterized enzymes. This method enables cellular validation of the specificity of small molecule DUB inhibitors. GENERAL SIGNIFICANCE: Molecular probes combined with mass spectrometry-based proteomics and cellular assays represent a powerful approach for discovery and functional validation, a concept that can be expanded to other enzyme classes. This addresses a need for more informative cell-based assays that are required to accelerate the drug development process. This article is part of a Special Issue entitled: Ubiquitin Drug Discovery and Diagnostics. |
| spellingShingle | Kramer, H Nicholson, B Kessler, B Altun, M Detection of ubiquitin-proteasome enzymatic activities in cells: application of activity-based probes to inhibitor development. |
| title | Detection of ubiquitin-proteasome enzymatic activities in cells: application of activity-based probes to inhibitor development. |
| title_full | Detection of ubiquitin-proteasome enzymatic activities in cells: application of activity-based probes to inhibitor development. |
| title_fullStr | Detection of ubiquitin-proteasome enzymatic activities in cells: application of activity-based probes to inhibitor development. |
| title_full_unstemmed | Detection of ubiquitin-proteasome enzymatic activities in cells: application of activity-based probes to inhibitor development. |
| title_short | Detection of ubiquitin-proteasome enzymatic activities in cells: application of activity-based probes to inhibitor development. |
| title_sort | detection of ubiquitin proteasome enzymatic activities in cells application of activity based probes to inhibitor development |
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