Silent Encoding of Chemical Post-Translational Modifications in Phage-Displayed Libraries.
In vitro selection of chemically modified peptide libraries presented on phage, while a powerful technology, is limited to one chemical post-translational modification (cPTM) per library. We use unique combinations of redundant codons to encode cPTMs with "silent barcodes" to trace multiple modifica...
| Publicado en: | Journal of the American Chemical Society Vol. 138; no. 1; pp. 32 - 36 |
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| Autores principales: | , , , , , , |
| Formato: | Artículo |
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American Chemical Society
1/13/2016
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| Materias: | |
| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=hlh&AN=113302198&site=ehost-live header: @attributes: shortDbName: hlh uiTerm: 113302198 longDbName: Humanities International Complete uiTag: AN controlInfo: bkinfo: jinfo: jid: 00027863 ACS jtl: Journal of the American Chemical Society issn: 00027863 maglogo: N pubinfo: dt: 1/13/2016 vid: 138 iid: 1 pid: 997 pub: American Chemical Society artinfo: ui: 113302198 10.1021/jacs.5b10390 ppf: 32 ppct: 4 formats: tig: atl: Silent Encoding of Chemical Post-Translational Modifications in Phage-Displayed Libraries. aug: au: Tjhung, Katrina F. Kitov, Pavel I. Ng, Simon Kitova, Elena N. Lu Deng Klassen, John S. Derda, Ratmir affil: Department of Chemistry, University of Alberta, Edmonton, Alberta, Canada T6G 2G su: Bacteriophages Microbial peptides Genetic code Carbonic anhydrase Sulfonamides sug: subj: Bacteriophages Microbial peptides Genetic code Carbonic anhydrase Sulfonamides ab: In vitro selection of chemically modified peptide libraries presented on phage, while a powerful technology, is limited to one chemical post-translational modification (cPTM) per library. We use unique combinations of redundant codons to encode cPTMs with "silent barcodes" to trace multiple modifications within a mixed modified library. As a proof of concept, we produced phage-displayed peptide libraries Ser-[X]-Gly-Gly-Gly, with Gly and Ser encoded using unique combinations of codons (TCA-[X]-GGAGGAGGA, AGT-[X]-GGTGGTGGT, etc., where [X] denotes a random NNK library). After separate chemical modification and pooling, mixed-modified libraries can be panned and deep-sequenced to identify the enriched peptide sequence and the accompanying cPTM simultaneously. We panned libraries bearing combinations of modifications (sulfonamide, biotin, mannose) against matched targets (carbonic anhydrase, streptavidin, concanavalin A) to identify desired ligands. Synthesis and validation of sequences identified by deep sequencing revealed that specific cPTMs are significantly enriched in panning against the specific targets. Panning on carbonic anhydrase yielded a potent ligand, sulfonamide-WIVP, with K = 6.7 ± 2.1 nM, a 20-fold improvement compared with the control ligand sulfonamide-GGGG. Silent encoding of multiple cPTMs can be readily incorporated into other in vitro display technologies such as bacteriophage T7 or mRNA display. pubtype: Academic Journal doctype: Article src: R language: English refInfo: copyright: @attributes: flag: Y dt: @attributes: year: 2016 holdings: @attributes: islocal: N |
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