Structural basis for transthiolation intermediates in the ubiquitin pathway Journal Article
| Authors: | Kochańczyk, T.; Hann, Z. S.; Lux, M. C.; Delos Reyes, A. M. V.; Ji, C.; Tan, D. S.; Lima, C. D. |
| Article Title: | Structural basis for transthiolation intermediates in the ubiquitin pathway |
| Abstract: | Transthiolation (also known as transthioesterification) reactions are used in the biosynthesis of acetyl coenzyme A, fatty acids and polyketides, and for post-translational modification by ubiquitin (Ub) and ubiquitin-like (Ubl) proteins1–3. For the Ub pathway, E1 enzymes catalyse transthiolation from an E1~Ub thioester to an E2~Ub thioester. Transthiolation is also required for transfer of Ub from an E2~Ub thioester to HECT (homologous to E6AP C terminus) and RBR (ring-between-ring) E3 ligases to form E3~Ub thioesters4–6. How isoenergetic transfer of thioester bonds is driven forward by enzymes in the Ub pathway remains unclear. Here we isolate mimics of transient transthiolation intermediates for E1–Ub(T)–E2 and E2–Ub(T)–E3HECT complexes (where T denotes Ub in a thioester or Ub undergoing transthiolation) using a chemical strategy with native enzymes and near-native Ub to capture and visualize a continuum of structures determined by single-particle cryo-electron microscopy. These structures and accompanying biochemical experiments illuminate conformational changes in Ub, E1, E2 and E3 that are coordinated with the chemical reactions to facilitate directional transfer of Ub from each enzyme to the next. © The Author(s) 2024. |
| Keywords: | controlled study; ubiquitin; mass spectrometry; electron microscopy; metabolism; ubiquitin protein ligase; carboxy terminal sequence; protein degradation; protein; protein interaction; enzyme activity; chemistry; protein processing; protein processing, post-translational; sexual orientation; crystal structure; models, molecular; esterification; conformational transition; crystallization; site directed mutagenesis; ubiquitin-activating enzymes; ubiquitin-conjugating enzymes; fatty acid; ubiquitin-protein ligases; catalyst; molecular model; image reconstruction; polyacrylamide gel electrophoresis; enzyme active site; schizosaccharomyces pombe; experimental study; ubiquitin conjugating enzyme; cryoelectron microscopy; affinity chromatography; electrospray mass spectrometry; liquid chromatography-mass spectrometry; ovalbumin; chemical method; polyketide; sanger sequencing; human; article; ultra performance liquid chromatography |
| Journal Title: | Nature |
| Volume: | 633 |
| Issue: | 8028 |
| ISSN: | 0028-0836 |
| Publisher: | Nature Publishing Group |
| Date Published: | 2024-09-05 |
| Start Page: | 216 |
| End Page: | 223 |
| Language: | English |
| DOI: | 10.1038/s41586-024-07828-9 |
| PUBMED: | 39143218 |
| PROVIDER: | scopus |
| PMCID: | PMC11374688 |
| DOI/URL: | |
| Notes: | Article -- MSK Cancer Center Support Grant (P30 CA008748) acknowledged in PubMed and PDF -- MSK corresponding authors are Derek Tan and Christopher Lima -- Source: Scopus |
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