4ZPZ

Crystal Structure of Semi-synthetic Ubiquitin with Phospho-Ser65 and Ala46Cys

Summary for 4ZPZ

• Entry DOI: 10.2210/pdb4zpz/pdb
• Descriptor: Polyubiquitin-B (2 entities in total)
• Functional Keywords: chromosomal protein, cell signalling, signaling protein
• Biological source: Homo sapiens (Human)
• Cellular location: Ubiquitin: Cytoplasm : P0CG47
• Total number of polymer chains: 2
• Total formula weight: 17377.75

Authors

Han, C.,Virdee, S. (deposition date: 2015-05-08, release date: 2015-06-10, Last modification date: 2024-11-13)

Primary citation

Han, C.,Pao, K.C.,Kazlauskaite, A.,Muqit, M.M.,Virdee, S.
A Versatile Strategy for the Semisynthetic Production of Ser65 Phosphorylated Ubiquitin and Its Biochemical and Structural Characterisation.
Chembiochem, 16:1574-1579, 2015

PubMed Abstract: 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.

PubMed: 26010437
DOI: 10.1002/cbic.201500185

Experimental method

X-RAY DIFFRACTION (1.54 Å)