Active site remodelling accompanies thioester bond formation in the SUMO E1 Journal Article
| Authors: | Olsen, S. K.; Capili, A. D.; Lu, X.; Tan, D. S.; Lima, C. D. |
| Article Title: | Active site remodelling accompanies thioester bond formation in the SUMO E1 |
| Abstract: | E1 enzymes activate ubiquitin (Ub) and ubiquitin-like (Ubl) proteins in two steps by carboxy-terminal adenylation and thioester bond formation to a conserved catalytic cysteine in the E1 Cys domain. The structural basis for these intermediates remains unknown. Here we report crystal structures for human SUMO E1 in complex with SUMO adenylate and tetrahedral intermediate analogues at 2.45 and 2.6 Ã.., respectively. These structures show that side chain contacts to ATPÂ •Mg are released after adenylation to facilitate a 130 degree rotation of the Cys domain during thioester bond formation that is accompanied by remodelling of key structural elements including the helix that contains the E1 catalytic cysteine, the crossover and re-entry loops, and refolding of two helices that are required for adenylation. These changes displace side chains required for adenylation with side chains required for thioester bond formation. Mutational and biochemical analyses indicate these mechanisms are conserved in other E1s. © 2010 Macmillan Publishers Limited. All rights reserved. |
| Keywords: | controlled study; ubiquitin; chemical analysis; protein conformation; enzyme activity; mutational analysis; amino acid sequence; conserved sequence; molecular sequence data; protein synthesis; saccharomyces cerevisiae; crystal structure; models, molecular; crystallography, x-ray; saccharomyces cerevisiae proteins; adenosine triphosphate; protein structure; sumo-1 protein; ubiquitin-activating enzymes; catalytic domain; catalyst; cysteine; thioester; biocatalysis; small ubiquitin-related modifier proteins; chemical bond; sulfides; ubiquitins; adenosine phosphate; adenylation; magnesium; ester; modeling; sumo 1 protein |
| Journal Title: | Nature |
| Volume: | 463 |
| Issue: | 7283 |
| ISSN: | 0028-0836 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2010-02-18 |
| Start Page: | 906 |
| End Page: | 912 |
| Language: | English |
| DOI: | 10.1038/nature08765 |
| PUBMED: | 20164921 |
| PROVIDER: | scopus |
| PMCID: | PMC2866016 |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 8" - "Export Date: 20 April 2011" - "CODEN: NATUA" - "Source: Scopus" |
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