Structural Basis for the Inhibitory Effects of Ubistatins in the Ubiquitin-Proteasome Pathway.
The discovery of ubistatins, small molecules that impair proteasomal degradation of proteins by directly binding to polyubiquitin, makes ubiquitin itself a potential therapeutic target. Although ubistatins have the potential for drug development and clinical applications, the lack of structural details of ubiquitin-ubistatin interactions has impeded their development. Here, we characterized a panel of new ubistatin derivatives using functional and binding assays. The structures of ubiquitin complexes with ubistatin B and hemi-ubistatin revealed direct interactions with ubiquitin's hydrophobic surface patch and the basic/polar residues surrounding it. Ubistatin B binds ubiquitin and diubiquitin tighter than a high-affinity ubiquitin receptor and shows strong preference for K48 linkages over K11 and K63. Furthermore, ubistatin B shields ubiquitin conjugates from disassembly by a range of deubiquitinases and by the 26S proteasome. Finally, ubistatin B penetrates cancer cells and alters the cellular ubiquitin landscape. These findings highlight versatile properties of ubistatins and have implications for their future development and use in targeting ubiquitin-signaling pathways.
Pubmed ID: 29153505
Research resources used in this publication
- TopSpin (RRID:SCR_014227) (software resource)
- PyMOL (RRID:SCR_000305) (software resource)
- Sparky (RRID:SCR_014228) (software resource)
- Fiji (RRID:SCR_002285) (software resource)
- Sparky (RRID:SCR_014228) (software resource)
- TopSpin (RRID:SCR_014227) (software resource)
- PyMOL (RRID:SCR_000305) (software resource)
- Fiji (RRID:SCR_002285) (software resource)
Additional research tools detected in this publication
Antibodies used in this publication
- Goat anti-Rabbit IgG H&L (Cy5 ®) secondary antibody (RRID:AB_10679461)
- Goat Anti-Rabbit IgG (H + L)-HRP Conjugate (RRID:AB_11125142)
- Mouse Anti-HA Monoclonal Antibody, Unconjugated, Clone 12CA5 (RRID:AB_514505)
- ubiquitin (RRID:AB_2315524)
- Rabbit Anti-Ubiquitin, Lys63-Specific Monoclonal antibody, Unconjugated, Clone apu3 (RRID:AB_1587580)
- Rabbit Anti-Ubiquitin, Lys48-Specific Monoclonal antibody, Unconjugated, Clone apu2 (RRID:AB_1587578)
- Anti-Ubiquitin K11 linkage Antibody, clone 2A3/2E6 (RRID:AB_2713901)
Associated grants
- Agency: NIGMS NIH HHS, United States
Id: R01 GM095755 - Agency: NIDDK NIH HHS, United States
Id: P30 DK079307 - Agency: NIGMS NIH HHS, United States
Id: R01 GM075061 - Agency: NIGMS NIH HHS, United States
Id: R01 GM065334 - Agency: NCI NIH HHS, United States
Id: N01CO12400
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This is a list of tools and resources that we have found mentioned in this publication.
ChemAxon (tool)
RRID:SCR_004111
Commercial organization that provides cheminformatics software platforms, applications and services to optimize the value of chemistry information in life science and other R&D. This software enables structure visualization and management, property predictions and calculations, virtual synthesis, screening, clustering and drug design.
View all literature mentionsGoat anti-Rabbit IgG H&L (Cy5 ®) secondary antibody (antibody)
RRID:AB_10679461
This polyclonal targets Goat anti-Rabbit IgG H&L (Cy5 ®) secondary antibody
View all literature mentionsGoat Anti-Rabbit IgG (H + L)-HRP Conjugate (antibody)
RRID:AB_11125142
This polyclonal secondary targets IgG (H+L)
View all literature mentionsMouse Anti-HA Monoclonal Antibody, Unconjugated, Clone 12CA5 (antibody)
RRID:AB_514505
This monoclonal targets HA
View all literature mentionsRabbit Anti-Ubiquitin, Lys63-Specific Monoclonal antibody, Unconjugated, Clone apu3 (antibody)
RRID:AB_1587580
This monoclonal targets Ubiquitin, Lys63-Specific
View all literature mentionsRabbit Anti-Ubiquitin, Lys48-Specific Monoclonal antibody, Unconjugated, Clone apu2 (antibody)
RRID:AB_1587578
This monoclonal targets Ubiquitin, Lys48-Specific
View all literature mentionsAnti-Ubiquitin K11 linkage Antibody, clone 2A3/2E6 (antibody)
RRID:AB_2713901
This monoclonal targets Ubiquitin K11 linkage
View all literature mentionsTopSpin (software resource)
RRID:SCR_014227
Software package for NMR spectra acquisition, processing, and data analysis. Features include: deconvolution/spectrum simulation/iteration; comprehensive functionalities for processing, displaying and analyzing one and multi-dimensional spectra; and user customization. A full list of features is available on the website.
View all literature mentionsPyMOL (software resource)
RRID:SCR_000305
A user-sponsored molecular visualization software system on an open-source foundation. The software has the capabilities to view, render, animate, export, present and develop three dimensional molecular structures.
View all literature mentionsSparky (software resource)
RRID:SCR_014228
A graphical NMR assignment and integration program for proteins, nucleic acids, and other polymers. Sparky displays NMR spectra, the peaks of which users may pick, assign, and integrate using a graphical interface. Users can work with any number of 2, 3 or 4 dimensional spectra simultaneously. Spectra for input to Sparky can be produced with processing programs NMRPipe, Felix, VNMR, XWinNMR or UXNMR. Output consists of text peak lists showing assignments, chemical shifts, volumes, line widths, etc.
View all literature mentionsFiji (software resource)
RRID:SCR_002285
Software package as distribution of ImageJ and ImageJ2 together with Java, Java3D and plugins organized into coherent menu structure. Used to assist research in life sciences.
View all literature mentionsSparky (software resource)
RRID:SCR_014228
A graphical NMR assignment and integration program for proteins, nucleic acids, and other polymers. Sparky displays NMR spectra, the peaks of which users may pick, assign, and integrate using a graphical interface. Users can work with any number of 2, 3 or 4 dimensional spectra simultaneously. Spectra for input to Sparky can be produced with processing programs NMRPipe, Felix, VNMR, XWinNMR or UXNMR. Output consists of text peak lists showing assignments, chemical shifts, volumes, line widths, etc.
View all literature mentionsTopSpin (software resource)
RRID:SCR_014227
Software package for NMR spectra acquisition, processing, and data analysis. Features include: deconvolution/spectrum simulation/iteration; comprehensive functionalities for processing, displaying and analyzing one and multi-dimensional spectra; and user customization. A full list of features is available on the website.
View all literature mentionsPyMOL (software resource)
RRID:SCR_000305
A user-sponsored molecular visualization software system on an open-source foundation. The software has the capabilities to view, render, animate, export, present and develop three dimensional molecular structures.
View all literature mentionsFiji (software resource)
RRID:SCR_002285
Software package as distribution of ImageJ and ImageJ2 together with Java, Java3D and plugins organized into coherent menu structure. Used to assist research in life sciences.
View all literature mentions
