Autor: |
Cong, Han, Kuan-Chuan, Pao, Agne, Kazlauskaite, Miratul M K, Muqit, Satpal, Virdee |
Rok vydání: |
2015 |
Předmět: |
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Zdroj: |
Chembiochem |
ISSN: |
1439-7633 |
Popis: |
Ubiquitin phosphorylation is emerging as an important regulatory layer in the ubiquitin system. This is exemplified by the phosphorylation of ubiquitin on Ser65 by the Parkinson's disease-associated kinase PINK1, which mediates the activation of the E3 ligase Parkin. Additional phosphorylation sites on ubiquitin might also have important cellular roles. Here we report a versatile strategy for preparing phosphorylated ubiquitin. We biochemically and structurally characterise semisynthetic phospho-Ser65-ubiquitin. Unexpectedly, we observed disulfide bond formation between ubiquitin molecules, and hence a novel crystal form. The method outlined provides a direct approach to study the combinatorial effects of phosphorylation on ubiquitin function. Our analysis also suggests that disulfide engineering of ubiquitin could be a useful strategy for obtaining alternative crystal forms of ubiquitin species thereby facilitating structural validation. |
Databáze: |
OpenAIRE |
Externí odkaz: |
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