Profiling the Cross Reactivity of Ubiquitin with the Nedd8 Activating Enzyme by Phage Display
| Autor: | Jun Yin, Hermann Schindelin, Eric B. Villhauer, Keya Zhang, Hiroaki Kiyokawa, Karan Bhuripanyo, Ning Zheng, Heng Li, Chan Hee J. Choi, Bo Zhao |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Models
Molecular Proteomics Phage display Proteome Protein Conformation Ubiquitin-activating enzyme lcsh:Medicine Ubiquitin-Activating Enzymes Ubiquitin-conjugating enzyme Protein Engineering Biochemistry NEDD8 Ligases Protein neddylation Ubiquitin Drug Discovery NEDD8 Activating Enzyme Biomacromolecule-Ligand Interactions lcsh:Science 0303 health sciences Multidisciplinary Enzyme Classes 030302 biochemistry & molecular biology Conjugated Proteins Recombinant Proteins Enzymes Hydrophobic and Hydrophilic Interactions Cullin Protein Binding Research Article Protein Structure Molecular Sequence Data Biophysics Biology Protein Chemistry Enzyme Regulation 03 medical and health sciences Peptide Library Transferases Chemical Biology Synthetic Peptide Amino Acid Sequence ddc:610 Protein Interactions 030304 developmental biology Enzyme Kinetics lcsh:R Proteins Regulatory Proteins Mutation Ubiquitin-Conjugating Enzymes Enzyme Structure Biocatalysis biology.protein lcsh:Q Peptidomimetics |
| Zdroj: | PLoS ONE, Vol 8, Iss 8, p e70312 (2013) PLoS ONE |
| Popis: | The C-terminal peptides of ubiquitin (UB) and UB-like proteins (UBLs) play a key role in their recognition by the specific activating enzymes (E1s) to launch their transfer through the respective enzymatic cascades thus modifying cellular proteins. UB and Nedd8, a UBL regulating the activity of cullin-RING UB ligases, only differ by one residue at their C-termini; yet each has its specific E1 for the activation reaction. It has been reported recently that UAE can cross react with Nedd8 to enable its passage through the UB transfer cascade for protein neddylation. To elucidate differences in UB recognition by UAE and NAE, we carried out phage selection of a UB library with randomized C-terminal sequences based on the catalytic formation of UB similar to NAE thioester conjugates. Our results confirmed the previous finding that residue 72 of UB plays a "gate-keeping" role in E1 selectivity. We also found that diverse sequences flanking residue 72 at the UB C-terminus can be accommodated by NAE for activation. Furthermore heptameric peptides derived from the C-terminal sequences of UB variants selected for NAE activation can function as mimics of Nedd8 to form thioester conjugates with NAE and the downstream E2 enzyme Ubc12 in the Nedd8 transfer cascade. Once the peptides are charged onto the cascade enzymes, the full-length Nedd8 protein is effectively blocked from passing through the cascade for the critical modification of cullin. We have thus identified a new class of inhibitors of protein neddylation based on the profiles of the UB C-terminal sequences recognized by NAE. |
| Databáze: | OpenAIRE |
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