Work

Genetically Encoded Crosslinking Enables Identification of Multivalent Ubiquitin‐Deubiquitylating Enzyme Interactions

Year: 2023

Type: article

Abstract: Ubiquitin (Ub) proteoforms control nearly every aspect of eukaryotic cell biology through their diversity. Inspired by the widely used Ub C-terminal electrophiles (Ub-E), here we report the identification of multivalent binding of Ub with deubiquitylating enzymes (Dubs) using genetic code expansion (GCE) and crosslinking mass spectrometry. While the Ub-Es only gather structural information with the S1 Dub sites, we demonstrate that GCE of Ub with p-benzoyl-L-phenylalanine enables identification of interaction modes beyond the S1 site with a panel of Dubs of both eukaryotic and prokaryotic origin. Collectively, this represents the next generation of Ub-based affinity probes with a unique ability to unravel Ub interaction landscapes beyond what is afforded by cysteine-based chemistries.

Source: ChemBioChem

DOI: 10.1002/cbic.202300305

Authors: Rishi Patel, Kristos Negrón Terón, Mowei Zhou, Ernesto Nakayasu, Bryon Drown, Chittaranjan Das

Institutions: Purdue University West Lafayette, Pacific Northwest National Laboratory, Environmental Molecular Sciences Laboratory

Language: English

Topic: Ubiquitin and proteasome pathways

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $4,020

Funders: U.S. Department of Energy, Purdue University, National Cancer Institute, National Institute of General Medical Sciences