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Protein semisynthesis reveals plasticity in HECT E3 ubiquitin ligase mechanisms
Jiang, Hanjie ; Miller, Bryant D ; Viennet, Thibault , et al. Nat. Chem.,2024,16,1894-1905. DOI: 10.1038/s41557-024-01576-z PubMed ID: 39030419
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Abstract: Lys ubiquitination is catalysed by E3 ubiquitin ligases and is central to the regulation of protein stability and cell signalling in normal and disease states. There are gaps in our understanding of E3 mechanisms, and here we use protein semisynthesis, chemical rescue, microscale thermophoresis and other biochemical approaches to dissect the role of catalytic base/acid function and conformational interconversion in HECT-domain E3 catalysis. We demonstrate that there is plasticity in the use of the terminal side chain or backbone carboxylate for proton transfer in HECT E3 ubiquitin ligases reactions, with yeast Rsp5 orthologues appearing to be possible evolutionary intermediates. We also show that the HECT-domain ubiquitin covalent intermediate appears to eject the E2 conjugating enzyme, promoting catalytic turnover. These fndings provide key mechanistic insights into how protein ubiquitination occurs and provide a framework for understanding E3 functions and regulation.
Purchased from AmBeed: 84624-17-9 ; 121343-82-6 ; 136083-57-3 ; 1007840-62-1
CAS No. : | 84624-17-9 | MDL No. : | MFCD00062953 |
Formula : | C20H21NO4 | Boiling Point : | No data available |
Linear Structure Formula : | - | InChI Key : | UGNIYGNGCNXHTR-GOSISDBHSA-N |
M.W : | 339.39 | Pubchem ID : | 1549479 |
Synonyms : |
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Signal Word: | Warning | Class: | |
Precautionary Statements: | P261-P280-P305+P351+P338 | UN#: | |
Hazard Statements: | H302-H315-H319-H332-H335 | Packing Group: | |
GHS Pictogram: |
* All experimental methods are cited from the reference, please refer to the original source for details. We do not guarantee the accuracy of the content in the reference.