PDBsum entry 4zpz

Signaling protein

PDB id: 4zpz

Contents

Protein chains

72 a.a.

Waters ×145

PDB id:

4zpz

Name:

Signaling protein

Title:

Crystal structure of semi-synthetic ubiquitin with phospho-ser65 and ala46cys

Structure:

Polyubiquitin-b. Chain: a, b. Fragment: ubiquitin, unp residues 1-73. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: ubb. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

1.54Å R-factor: 0.140 R-free: 0.173

Authors:

C.Han,S.Virdee

Key ref:

C.Han et al. (2015). A Versatile Strategy for the Semisynthetic Production of Ser65 Phosphorylated Ubiquitin and Its Biochemical and Structural Characterisation. Chembiochem, 16, 1574-1579. PubMed id: 26010437 DOI: 10.1002/cbic.201500185

Date:

08-May-15 Release date: 10-Jun-15

Protein chains

P0CG47  (UBB_HUMAN) -  Polyubiquitin-B from Homo sapiens

Seq: 229 a.a.

Struc: 72 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

DOI no: 10.1002/cbic.201500185

Chembiochem 16:1574-1579 (2015)

PubMed id: 26010437

A Versatile Strategy for the Semisynthetic Production of Ser65 Phosphorylated Ubiquitin and Its Biochemical and Structural Characterisation.

C.Han, K.C.Pao, A.Kazlauskaite, M.M.Muqit, S.Virdee.

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.