Designed Semisynthetic Protein Inhibitors of Ub/Ubl E1 Activating Enzymes

Semisynthetic, mechanism-based protein inhibitors of ubiquitin (Ub) and ubiquitin-like modifier (Ubl) activating enzymes (E1s) have been developed to target E1-catalyzed adenylation and thioesterification of the Ub/Ubl C-terminus during the processes of protein SUMOylation and ubiquitination. The in...

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Bibliographic Details
Published in:Journal of the American Chemical Society 2010-02, Vol.132 (6), p.1748-1749
Main Authors: Lu, Xuequan, Olsen, Shaun K, Capili, Allan D, Cisar, Justin S, Lima, Christopher D, Tan, Derek S
Format: Article
Language:English
Subjects:
Drug Design
Enzyme Inhibitors - chemical synthesis
Enzyme Inhibitors - pharmacology
Models, Molecular
Protein Conformation
Substrate Specificity
SUMO-1 Protein - chemistry
SUMO-1 Protein - metabolism
Ubiquitin - chemistry
Ubiquitin - metabolism
Ubiquitin-Activating Enzymes - antagonists & inhibitors
Ubiquitin-Activating Enzymes - chemistry
Ubiquitin-Activating Enzymes - metabolism
Ubiquitins - chemistry
Ubiquitins - metabolism
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Summary:Semisynthetic, mechanism-based protein inhibitors of ubiquitin (Ub) and ubiquitin-like modifier (Ubl) activating enzymes (E1s) have been developed to target E1-catalyzed adenylation and thioesterification of the Ub/Ubl C-terminus during the processes of protein SUMOylation and ubiquitination. The inhibitors were generated by intein-mediated expressed protein ligation using a truncated Ub/Ubl protein (SUMO residues 1−94; Ub residues 1−71) with a C-terminal thioester and synthetic tripeptides having a C-terminal adenosine analogue and an N-terminal cysteine residue. SUMO-AMSN (4a) and Ub-AMSN (4b) contain a sulfamide group as a nonhydrolyzable mimic of the phosphate group in the cognate Ub/Ubl-AMP adenylate intermediate in the first half-reaction, and these constructs selectively inhibit SUMO E1 and Ub E1, respectively, in a dose-dependent manner. SUMO-AVSN (5a) and Ub-AVSN (5b) contain an electrophilic vinyl sulfonamide designed to trap the incoming E1 cysteine nucleophile (Uba2 Cys173 in SUMO E1; Uba1 Cys593 in Ub E1) in the second half-reaction, and these constructs selectively, covalently, and stably cross-link to SUMO E1 and Ub E1, respectively, in a cysteine nucleophile-dependent manner. These inhibitors are powerful tools to probe outstanding mechanistic questions in E1 function and can also be used to study the biological functions of E1 enzymes.
ISSN:0002-7863
1520-5126
DOI:10.1021/ja9088549