Genetically encoding phosphotyrosine and its nonhydrolyzable analog in bacteria
dc.creator | Luo, X | |
dc.creator | Fu, G | |
dc.creator | Wang, RE | |
dc.creator | Zhu, X | |
dc.creator | Zambaldo, C | |
dc.creator | Liu, R | |
dc.creator | Liu, T | |
dc.creator | Lyu, X | |
dc.creator | Du, J | |
dc.creator | Xuan, W | |
dc.creator | Yao, A | |
dc.creator | Reed, SA | |
dc.creator | Kang, M | |
dc.creator | Zhang, Y | |
dc.creator | Guo, H | |
dc.creator | Huang, C | |
dc.creator | Yang, PY | |
dc.creator | Wilson, IA | |
dc.creator | Schultz, PG | |
dc.creator | Wang, F | |
dc.date.accessioned | 2021-01-22T20:41:45Z | |
dc.date.available | 2021-01-22T20:41:45Z | |
dc.date.issued | 2017-08-01 | |
dc.identifier.issn | 1552-4450 | |
dc.identifier.issn | 1552-4469 | |
dc.identifier.doi | http://dx.doi.org/10.34944/dspace/4865 | |
dc.identifier.other | 28604693 (pubmed) | |
dc.identifier.uri | http://hdl.handle.net/20.500.12613/4883 | |
dc.description.abstract | © 2017 Nature America, Inc., part of Springer Nature. All rights reserved. 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. | |
dc.format.extent | 845-849 | |
dc.language.iso | en | |
dc.relation.haspart | Nature Chemical Biology | |
dc.relation.isreferencedby | Springer Science and Business Media LLC | |
dc.subject | Codon, Nonsense | |
dc.subject | Crystallography, X-Ray | |
dc.subject | Escherichia coli | |
dc.subject | Ligases | |
dc.subject | Models, Molecular | |
dc.subject | Molecular Structure | |
dc.subject | Phosphorylation | |
dc.subject | Phosphotyrosine | |
dc.subject | Recombinant Proteins | |
dc.title | Genetically encoding phosphotyrosine and its nonhydrolyzable analog in bacteria | |
dc.type | Article | |
dc.type.genre | Post-print | |
dc.relation.doi | 10.1038/nchembio.2405 | |
dc.ada.note | For Americans with Disabilities Act (ADA) accommodation, including help with reading this content, please contact scholarshare@temple.edu | |
dc.creator.orcid | Wang, Rongsheng|0000-0002-5749-7447 | |
dc.date.updated | 2021-01-22T20:41:41Z | |
refterms.dateFOA | 2021-01-22T20:41:45Z |