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dc.creatorLuo, X
dc.creatorFu, G
dc.creatorWang, RE
dc.creatorZhu, X
dc.creatorZambaldo, C
dc.creatorLiu, R
dc.creatorLiu, T
dc.creatorLyu, X
dc.creatorDu, J
dc.creatorXuan, W
dc.creatorYao, A
dc.creatorReed, SA
dc.creatorKang, M
dc.creatorZhang, Y
dc.creatorGuo, H
dc.creatorHuang, C
dc.creatorYang, PY
dc.creatorWilson, IA
dc.creatorSchultz, PG
dc.creatorWang, F
dc.date.accessioned2021-01-22T20:41:45Z
dc.date.available2021-01-22T20:41:45Z
dc.date.issued2017-08-01
dc.identifier.issn1552-4450
dc.identifier.issn1552-4469
dc.identifier.doihttp://dx.doi.org/10.34944/dspace/4865
dc.identifier.other28604693 (pubmed)
dc.identifier.urihttp://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.extent845-849
dc.language.isoen
dc.relation.haspartNature Chemical Biology
dc.relation.isreferencedbySpringer Science and Business Media LLC
dc.subjectCodon, Nonsense
dc.subjectCrystallography, X-Ray
dc.subjectEscherichia coli
dc.subjectLigases
dc.subjectModels, Molecular
dc.subjectMolecular Structure
dc.subjectPhosphorylation
dc.subjectPhosphotyrosine
dc.subjectRecombinant Proteins
dc.titleGenetically encoding phosphotyrosine and its nonhydrolyzable analog in bacteria
dc.typeArticle
dc.type.genrePost-print
dc.relation.doi10.1038/nchembio.2405
dc.ada.noteFor Americans with Disabilities Act (ADA) accommodation, including help with reading this content, please contact scholarshare@temple.edu
dc.creator.orcidWang, Rongsheng|0000-0002-5749-7447
dc.date.updated2021-01-22T20:41:41Z
refterms.dateFOA2021-01-22T20:41:45Z


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