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A native chemical ligation handle that enables the synthesis of advanced activity-based probes: diubiquitin as a case study.

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.

SUBMITTER: Mulder MP

PROVIDER: S-EPMC4159580 | biostudies-literature | 2014 May

REPOSITORIES: biostudies-literature

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A native chemical ligation handle that enables the synthesis of advanced activity-based probes: diubiquitin as a case study.

Mulder Monique P C MP El Oualid Farid F ter Beek Jarno J Ovaa Huib H

Chembiochem : a European journal of chemical biology 20140312 7

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 o ...[more]

PMID: 24623714

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