Work

Approaches to Evaluate the Impact of a Small‐Molecule Binder to a Noncatalytic Site of the Proteasome

Year: 2021

Type: article

Abstract: Proteasome activity is crucial for cell survival and proliferation. In recent years, small molecules have been discovered that can affect the catalytic activity of the proteasome. Rather than targeting the active sites of the proteasome, it might be possible to affect ubiquitin-dependent degradation of proteins by limiting the association of the 19S regulatory particle (19S RP) with the 20S core particle (20S CP) of the proteasome. We recently described the discovery of TXS-8, a peptoid that binds to Rpn-6. Rpn-6 is a proteasome-associated protein that makes critical contacts with the 19S RP and the 20S CP. Herein, we present a general workflow to evaluate the impact of a small-molecule binder on proteasome activity by using TXS-8 as an example. This workflow contains three steps in which specific probes or overexpressed proteins in cells are used to determine whether the hydrolysis activity of the proteasome is affected. Although, in our case, TXS-8 did not affect proteasome activity, our workflow is highly amenable to studying a variety of small-molecule-proteasome subunit interactions.

Source: ChemBioChem

DOI: 10.1002/cbic.202100023

Authors: Wenzhi Tian, Marianne E. Maresh, Darci J. Trader

Institution: Purdue University West Lafayette

Language: English

Topic: Ubiquitin and proteasome pathways

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: green

Funders: Purdue University Center for Cancer Research, National Cancer Institute, National Institute of General Medical Sciences, National Institute of Allergy and Infectious Diseases

Awards: G272352002, Cancer Center Support Grant, G5657832250, Development of Activity-Based Chemical Reporters to Differentiate Proteasome Isoforms in Cells