Protein degradation profile reveals dynamic nature of 20S proteasome small molecule stimulation

Coleman, Robert W.; Mohallem, Rodrigo; Aryal, Uma K.; Trader, Darci J.

doi:10.1039/d0cb00191k

Cited by 15 publications

(17 citation statements)

References 34 publications

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“…The pair (4, 12) however shows a slight decrease in stimulation. Across the series in Table 2, adding a larger substitution at the para position, such as methoxy (11) and t-butyl (13), or a nonpolar substitution, such as a methyl (10), trifluoromethyl (14), decreases 20S CP stimulation. This pattern is also evident in Table 1.…”

Section: Resultsmentioning

confidence: 99%

“…1B, and evaluated their ability to increase the ability of the 20S CP to degrade a fluorescent peptide reporter and alpha-synuclein, an intrinsically disordered protein that has been associated with Parkinson's disease. 4,10–12 When compared with AM-404, our best molecule, 17 , exhibited a similar in cellulo stimulatory activity of the 20S CP, as well as toxicity profile.…”

Section: Introductionmentioning

confidence: 92%

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Oleic amide derivatives as small molecule stimulators of the human proteasome's core particle

et al. 2022

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 92%

Oleic amide derivatives as small molecule stimulators of the human proteasome's core particle

et al. 2022

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“…We next aimed to test ( 3 ) for its ability to detect changes in proteasome activity in cells in the presence of proteasome stimulators or inhibitors. For the activity assays with modulators of the proteasome, we selected a set of previously established modulators of the multi‐subunit complex in both biochemical and cellular environments [31–34] . Three activators and two inhibitors were used at two different concentrations for our studies, Figure 5A.…”

Section: Resultsmentioning

confidence: 99%

“…For the activity assays with modulators of the proteasome, we selected a set of previously established modulators of the multi-subunit complex in both biochemical and cellular environments. [31][32][33][34] Three activators and two inhibitors were used at two different concentrations for our studies, Figure 5A. Cells were pre-treated with the small molecule inhibitor/stimulator for one hour, then (3) was added at 500 nM, and the cells were incubated for an additional hour.…”

Section: Proteasome Activity Assays Of Positive and Negative Modulato...mentioning

confidence: 99%

Solid‐Phase Synthesis and Application of a Clickable Version of Epoxomicin for Proteasome Activity Analysis

et al. 2022

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Degradation of proteins by the proteasome is an essential cellular process and one that many wish to study in a variety of disease types. There are commercially available probes that can monitor proteasome activity in cells, but they typically contain common fluorophores that limit their simultaneous use with other activity-based probes. In order to exchange the fluorophore or incorporate an enrichment tag, the proteasome probe likely has to be synthesized which can be cumbersome. Here, we describe a simple synthetic procedure that only requires one purification step to generate epoxomicin, a selective proteasome inhibitor, with a terminal alkyne. Through a copper-catalyzed cycloaddition, any moiety containing an azide can be incorporated into the probe. Many fluorophores are commercially available that contain an azide that can be "clicked", allowing this proteasome activity probe to be included into already established assays to monitor both proteasome activity and other cellular activities of interest.

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“…This holoenzyme consists of a barrel-shaped 20S core particle capped on one or both ends by 19S regulatory particles ( Figure 3B ; Budenholzer et al, 2017 ; Bard et al, 2018 ). Unfolded proteins are processed by subunits with proteolytic activities that reside in the 20S core particle ( Rennella et al, 2020 ; Coleman et al, 2021 ). The regulatory particle is responsible for substrate recognition, deubiquitination, and ATP-dependent substrate unfolding that enables subsequent translocation through the channel pore ( Lu et al, 2017 ).…”

Section: The Ub-proteasome System In Reversing Protein Aggregationmentioning

confidence: 99%

A Potential Mechanism for Targeting Aggregates With Proteasomes and Disaggregases in Liquid Droplets

Hayes

Sirvio

2022

Front. Aging Neurosci.

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Insoluble protein deposits are hallmarks of neurodegenerative disorders and common forms of dementia. The aberrant aggregation of misfolded proteins involves a complex cascade of events that occur over time, from the cellular to the clinical phase of neurodegeneration. Declining neuronal health through increased cell stress and loss of protein homeostasis (proteostasis) functions correlate with the accumulation of aggregates. On the cellular level, increasing evidence supports that misfolded proteins may undergo liquid-liquid phase separation (LLPS), which is emerging as an important process to drive protein aggregation. Studying, the reverse process of aggregate disassembly and degradation has only recently gained momentum, following reports of enzymes with distinct aggregate-disassembly activities. In this review, we will discuss how the ubiquitin-proteasome system and disaggregation machineries such as VCP/p97 and HSP70 system may disassemble and/or degrade protein aggregates. In addition to their canonically associated functions, these enzymes appear to share a common feature: reversibly assembling into liquid droplets in an LLPS-driven manner. We review the role of LLPS in enhancing the disassembly of aggregates through locally increasing the concentration of these enzymes and their co-proteins together within droplet structures. We propose that such activity may be achieved through the concerted actions of disaggregase machineries, the ubiquitin-proteasome system and their co-proteins, all of which are condensed within transient aggregate-associated droplets (TAADs), ultimately resulting in aggregate clearance. We further speculate that sustained engagement of these enzymatic activities within TAADs will be detrimental to normal cellular functions, where these activities are required. The possibility of facilitating endogenous disaggregation and degradation activities within TAADs potentially represents a novel target for therapeutic intervention to restore protein homeostasis at the early stages of neurodegeneration.

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Protein degradation profile reveals dynamic nature of 20S proteasome small molecule stimulation

Abstract: Small molecule stimulators of the 20S core particle of the proteasome can lead to the degradation of a variety of protein substrates.

Cited by 15 publications

References 34 publications

Oleic amide derivatives as small molecule stimulators of the human proteasome's core particle

Oleic amide derivatives as small molecule stimulators of the human proteasome's core particle

Solid‐Phase Synthesis and Application of a Clickable Version of Epoxomicin for Proteasome Activity Analysis

A Potential Mechanism for Targeting Aggregates With Proteasomes and Disaggregases in Liquid Droplets

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