Structural and Functional Characterization of Ubiquitin Variant Inhibitors of USP15
| Autor: | Jason Moffat, Frank W. Schmitges, Bradley B. Brasher, Carsten Schwerdtfeger, Patrick Jaynes, Michael F. Moran, Pieter Johan Adam Eichhorn, Alex U. Singer, Joan Teyra, G. Boehmelt, M.J. Polyak, Philippe Gros, J. Tong, Derek F. Ceccarelli, Nassima Fodil, Jonathan R. Krieger, S. Kit Leng Lui, M. Lenter, Frank Sicheri, Sachdev S. Sidhu |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Models
Molecular Protein Conformation alpha-Helical TRIM25 Ubiquitin-Protein Ligases Dimer Genetic Vectors Gene Expression Crystallography X-Ray Substrate Specificity Deubiquitinating enzyme Transforming Growth Factor beta1 Tripartite Motif Proteins 03 medical and health sciences chemistry.chemical_compound Ubiquitin Structural Biology Catalytic Domain Escherichia coli Humans Protein Interaction Domains and Motifs Amino Acid Sequence Cloning Molecular Molecular Biology 030304 developmental biology 0303 health sciences Sequence Homology Amino Acid biology Chemistry 030302 biochemistry & molecular biology Ubiquitination Active site Recombinant Proteins Cell biology HEK293 Cells biology.protein Phosphorylation Protein Conformation beta-Strand Ubiquitin-Specific Proteases Protein Multimerization Sequence Alignment Function (biology) Protein Binding Transcription Factors Deubiquitination |
| Zdroj: | Structure. 27:590-605.e5 |
| ISSN: | 0969-2126 |
| Popis: | Summary The multi-domain deubiquitinase USP15 regulates diverse eukaryotic processes and has been implicated in numerous diseases. We developed ubiquitin variants (UbVs) that targeted either the catalytic domain or each of three adaptor domains in USP15, including the N-terminal DUSP domain. We also designed a linear dimer (diUbV), which targeted the DUSP and catalytic domains, and exhibited enhanced specificity and more potent inhibition of catalytic activity than either UbV alone. In cells, the UbVs inhibited the deubiquitination of two USP15 substrates, SMURF2 and TRIM25, and the diUbV inhibited the effects of USP15 on the transforming growth factor β pathway. Structural analyses revealed that three distinct UbVs bound to the catalytic domain and locked the active site in a closed, inactive conformation, and one UbV formed an unusual strand-swapped dimer and bound two DUSP domains simultaneously. These inhibitors will enable the study of USP15 function in oncology, neurology, immunology, and inflammation. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|