Designed semisynthetic protein inhibitors of Ub/Ubl E1 activating enzymes Journal Article
| Authors: | Lu, X.; Olsen, S. K.; Capili, A. D.; Cisar, J. S.; Lima, C. D.; Tan, D. S. |
| Article Title: | Designed semisynthetic protein inhibitors of Ub/Ubl E1 activating enzymes |
| Abstract: | (Chemical Equation Presented) 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. Copyright © 2010 American Chemical Society. |
| Keywords: | unclassified drug; ubiquitin; protein conformation; protein; enzyme activity; drug design; ubiquitination; enzyme inhibitors; substrate specificity; models, molecular; esterification; chemical reactions; sumoylation; sumo-1 protein; ubiquitin-activating enzymes; chemical equations; enzymes; synthesis; sulfur compounds; protein inhibitor; ubiquitins; adenylate; enzyme; adenylation; thioesters; biological functions; c-terminus; dose-dependent manner; expressed protein ligation; n-terminals; phosphate group; protein inhibitors; thioesterification; tri-peptides; ubiquitin-like; catalysts; nucleophiles; reaction intermediates; e1 activating enzyme; ubiquitin like protein |
| Journal Title: | Journal of the American Chemical Society |
| Volume: | 132 |
| Issue: | 6 |
| ISSN: | 0002-7863 |
| Publisher: | American Chemical Society |
| Date Published: | 2010-02-17 |
| Start Page: | 1748 |
| End Page: | 1749 |
| Language: | English |
| DOI: | 10.1021/ja9088549 |
| PUBMED: | 20099854 |
| PROVIDER: | scopus |
| PMCID: | PMC2830896 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 3" - "Export Date: 20 April 2011" - "CODEN: JACSA" - "Source: Scopus" |
Altmetric
Citation Impact
BMJ Impact Analytics
Related MSK Work