1. A native chemical ligation handle that enables the synthesis of advanced activity-based probes: diubiquitin as a case study.
- Author
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Mulder MP, El Oualid F, ter Beek J, and Ovaa H
- Subjects
- Animals, Carbocyanines chemistry, Catalytic Domain, Cell Line, Tumor, Electrophoresis, Endopeptidases chemistry, Endopeptidases metabolism, Humans, Mice, Substrate Specificity, Fluorescent Dyes chemistry, Ubiquitins chemistry
- Abstract
We present the development of a native chemical ligation handle that also functions as a masked electrophile that can be liberated during synthesis when required. This handle can thus be used for the synthesis of complex activity-based probes. We describe the use of this handle in the generation of linkage-specific activity-based deubiquitylating enzyme probes that contain substrate context and closely mimic the native ubiquitin isopeptide linkage. We have generated activity-based probes based on all seven isopeptide-linked diubiquitin topoisomers and demonstrated their structural integrity and ability to label DUBs in a linkage-specific manner., (© 2014 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.)
- Published
- 2014
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