5U36

Crystal Structure Of A Mutant M. Jannashii Tyrosyl-tRNA Synthetase

Summary for 5U36

• Entry DOI: 10.2210/pdb5u36/pdb
• Related: 1J1U 1U7D
• Descriptor: Tyrosine--tRNA ligase (1 entity in total)
• Functional Keywords: tyrosine phosphorylation, ligase
• Biological source: Methanocaldococcus jannaschii (strain ATCC 43067 / DSM 2661 / JAL-1 / JCM 10045 / NBRC 100440)
• Total number of polymer chains: 2
• Total formula weight: 72011.58

Authors

Luo, X.,Fu, G.,Zhu, X.,Wilson, I.A.,Wang, F. (deposition date: 2016-12-01, release date: 2017-06-07, Last modification date: 2023-10-04)

Primary citation

Luo, X.,Fu, G.,Wang, R.E.,Zhu, X.,Zambaldo, C.,Liu, R.,Liu, T.,Lyu, X.,Du, J.,Xuan, W.,Yao, A.,Reed, S.A.,Kang, M.,Zhang, Y.,Guo, H.,Huang, C.,Yang, P.Y.,Wilson, I.A.,Schultz, P.G.,Wang, F.
Genetically encoding phosphotyrosine and its nonhydrolyzable analog in bacteria.
Nat. Chem. Biol., 13:845-849, 2017

PubMed Abstract: Tyrosine phosphorylation is a common protein post-translational modification that plays a critical role in signal transduction and the regulation of many cellular processes. Using a propeptide strategy to increase cellular uptake of O-phosphotyrosine (pTyr) and its nonhydrolyzable analog 4-phosphomethyl-L-phenylalanine (Pmp), we identified an orthogonal aminoacyl-tRNA synthetase-tRNA pair that allows site-specific incorporation of both pTyr and Pmp into recombinant proteins in response to the amber stop codon in Escherichia coli in good yields. The X-ray structure of the synthetase reveals a reconfigured substrate-binding site, formed by nonconservative mutations and substantial local structural perturbations. We demonstrate the utility of this method by introducing Pmp into a putative phosphorylation site and determining the affinities of the individual variants for the substrate 3BP2. In summary, this work provides a useful recombinant tool to dissect the biological functions of tyrosine phosphorylation at specific sites in the proteome.

PubMed: 28604693
DOI: 10.1038/nchembio.2405

Experimental method

X-RAY DIFFRACTION (3.03 Å)