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Ubiquitin Chain Analysis by Parallel Reaction Monitoring

Ubiquitin Chain Analysis by Parallel Reaction Monitoring

Published on: June 17, 2020

The Length of a Ubiquitin Chain: A General Factor for Selective Recognition by Ubiquitin-Binding Proteins

Joachim Lutz¹, Eva Höllmüller¹, Martin Scheffner¹

¹Departments of Chemistry and Biology, Konstanz Research School Chemical Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.

Angewandte Chemie (International Ed. in English)

|April 18, 2020

Abstract:

The attachment of ubiquitin (Ub) chains of various length to proteins is a prevalent posttranslational modification in eukaryotes. The fate of a modified protein is determined by Ub-binding proteins (UBPs), which interact with Ub chains in a linkage-selective manner. However, the impact and functional consequences of chain length on the binding selectivity of UBPs remain mostly elusive. We have generated Ub chains of defined length and linkage by using click chemistry and GELFrEE fractionation. These defined polymers were used in affinity-based enrichment assays to identify length- and linkage-selective interaction partners on a proteome-wide scale. For the first time, it is revealed that the length of a Ub chain generally has a major impact on its ability to be selectively recognized by UBPs.

Keywords:

Ub-binding proteins chain length posttranslational modification ubiquitin

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