Direct characterization of E2-dependent target specificity and processivity using an artificial p27-linker-E2 ubiquitination system
| Autor: | Young Ho Jeon, Junsang Ko, Hyun Jung Kim, Hae-Kap Cheong, Seong-Ock Kim, Chaejoon Cheong, Yun-Seok Choi, Byong-Seok Choi, Kyoung-Seok Ryu |
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| Rok vydání: | 2008 |
| Předmět: |
Recombinant Fusion Proteins
Ubiquitin-Protein Ligases In Vitro Techniques Direct transfer Ubiquitin-conjugating enzyme Models Biological Biochemistry Anaphase-Promoting Complex-Cyclosome Substrate Specificity Ubiquitin Proliferating Cell Nuclear Antigen Humans Molecular Biology biology Chemistry Lysine Intracellular Signaling Peptides and Proteins Ubiquitination Ubiquitin-Protein Ligase Complexes General Medicine Processivity Fusion protein Ubiquitin ligase Cell biology Ubiquitin-Conjugating Enzymes biology.protein Target protein Linker Cyclin-Dependent Kinase Inhibitor p27 |
| Zdroj: | BMB Reports. 41:852-857 |
| ISSN: | 1976-6696 |
| Popis: | Little attention has been paid to the specificity between E2 and the target protein during ubiquitination, although RING-E3 induces a potential intra-molecular reaction by mediating the direct transfer of ubiquitin from E2 to the target protein. We have constructed artificial E2 fusion proteins in which a target protein (p27) is tethered to one of six E2s via a flexible linker. Interestingly, only three E2s (UbcH5b, hHR6b, and Cdc34) are able to ubiquitinate p27 via an intra-molecular reaction in this system. Although the first ubiquitination of p27 (p27-Ub) by Cdc34 is less efficient than that of UbcH5b and hHR6b, the additional ubiquitin attachment to p27-Ub by Cdc34 is highly efficient. The E2 core of Cdc34 provides specificity to p27, and the residues 184-196 are required for possessive ubiquitination by Cdc34. We demonstrate direct E2 specificity for p27 and also show that differential ubiquitin linkages can be dependent on E2 alone. |
| Databáze: | OpenAIRE |
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