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. 2021:2365:283-300.
doi: 10.1007/978-1-0716-1665-9_15.

Profiling CELMoD-Mediated Degradation of Cereblon Neosubstrates

Joel W Thompson  1 , Thomas Clayton  1 , Gody Khambatta  1 , Leslie A Bateman  1 , Christopher W Carroll  1 , Philip P Chamberlain  1 , Mary E Matyskiela  2
Affiliations

Profiling CELMoD-Mediated Degradation of Cereblon Neosubstrates

Joel W Thompson et al. Methods Mol Biol. 2021.

Abstract

Targeted protein degradation is garnering increased attention as a therapeutic modality due in part to its promise of modulating targets previously considered undruggable. Cereblon E3 Ligase Modulating Drugs (CELMoDs) are one of the most well-characterized therapeutics employing this modality. CELMoDs hijack Cereblon E3 ligase activity causing neosubstrates to be ubiquitinated and degraded in the proteasome. Here, we describe a suite of assays-cellular substrate degradation, confirmation of CELMoD mechanism of action, in vitro ubiquitination, and Cereblon binding-that can be used to characterize CELMoD-mediated degradation of Cereblon neosubstrates. While the assays presented herein can be run independently, when combined they provide a strong platform to support the discovery and optimization of CELMoDs and fuel validation of targets degraded by this drug modality.

Keywords: CELMoD; Cellular neosubstrate degradation; Cereblon; Cereblon binding; In vitro ubiquitination; Targeted protein degradation.

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