FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W3036404411> ?p ?o ?g. }

• W3036404411 endingPage "4704" @default.
• W3036404411 startingPage "4690" @default.
• W3036404411 abstract "Genetic code expansion (GCE) technologies incorporate non-canonical amino acids (ncAAs) into proteins at amber stop codons. To avoid unwanted truncated protein and improve ncAA–protein yields, genomically recoded strains of Escherichia coli lacking Release Factor 1 (RF1) are becoming increasingly popular expression hosts for GCE applications. In the absence of RF1, however, endogenous near-cognate amber suppressing tRNAs can lead to contaminating protein forms with natural amino acids in place of the ncAA. Here, we show that a second-generation amino-acyl tRNA synthetase (aaRS)/tRNACUA pair for site-specific incorporation of 3-nitro-tyrosine could not outcompete near-cognate suppression in an RF1-deficient expression host and therefore could not produce homogenously nitrated protein. To resolve this, we used Rosetta to target positions in the nitroTyr aaRS active site for improved substrate binding, and then constructed of a small library of variants to subject to standard selection protocols. The top selected variant had an ~ 2-fold greater efficiency, and remarkably, this relatively small improvement enabled homogeneous incorporation of nitroTyr in an RF1-deficient expression host and thus eliminates truncation issues associated with typical RF1-containing expression hosts. Structural and biochemical data suggest the aaRS efficiency improvement is based on higher affinity substrate binding. Taken together, the modest improvement in aaRS efficiency provides a large practical impact and expands our ability to study the role protein nitration plays in disease development through producing homogenous, truncation-free nitroTyr-containing protein. This work establishes Rosetta-guided design and incremental aaRS improvement as a viable and accessible path to improve GCE systems challenged by truncation and/or near-cognate suppression issues." @default.
• W3036404411 created "2020-06-25" @default.
• W3036404411 creator A5015680529 @default.
• W3036404411 creator A5021167805 @default.
• W3036404411 creator A5026956570 @default.
• W3036404411 creator A5027346080 @default.
• W3036404411 creator A5030497987 @default.
• W3036404411 creator A5041427467 @default.
• W3036404411 creator A5058485400 @default.
• W3036404411 creator A5087060483 @default.
• W3036404411 creator A5091612311 @default.
• W3036404411 date "2020-07-01" @default.
• W3036404411 modified "2023-10-14" @default.
• W3036404411 title "Overcoming Near-Cognate Suppression in a Release Factor 1-Deficient Host with an Improved Nitro-Tyrosine tRNA Synthetase" @default.
• W3036404411 cites W1067029065 @default.
• W3036404411 cites W1216718361 @default.
• W3036404411 cites W1969449009 @default.
• W3036404411 cites W1975952514 @default.
• W3036404411 cites W1989760659 @default.
• W3036404411 cites W2044289704 @default.
• W3036404411 cites W2051947456 @default.
• W3036404411 cites W2053235167 @default.
• W3036404411 cites W2058079617 @default.
• W3036404411 cites W2077785805 @default.
• W3036404411 cites W2078290510 @default.
• W3036404411 cites W2086361102 @default.
• W3036404411 cites W2090855681 @default.
• W3036404411 cites W2144296712 @default.
• W3036404411 cites W2163266294 @default.
• W3036404411 cites W2166595842 @default.
• W3036404411 cites W2227591440 @default.
• W3036404411 cites W2261521410 @default.
• W3036404411 cites W2270583408 @default.
• W3036404411 cites W2327030163 @default.
• W3036404411 cites W2331636826 @default.
• W3036404411 cites W2341299591 @default.
• W3036404411 cites W2529122827 @default.
• W3036404411 cites W2734877409 @default.
• W3036404411 cites W2743989173 @default.
• W3036404411 cites W2752361102 @default.
• W3036404411 cites W2754403265 @default.
• W3036404411 cites W2801979949 @default.
• W3036404411 cites W2884133005 @default.
• W3036404411 cites W2891841439 @default.
• W3036404411 cites W2898867648 @default.
• W3036404411 cites W2944014204 @default.
• W3036404411 cites W2944969418 @default.
• W3036404411 cites W2947191811 @default.
• W3036404411 cites W2949014628 @default.
• W3036404411 cites W2959338423 @default.
• W3036404411 cites W2997259560 @default.
• W3036404411 cites W4252192583 @default.
• W3036404411 cites W946883613 @default.
• W3036404411 doi "https://doi.org/10.1016/j.jmb.2020.06.014" @default.
• W3036404411 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/7665880" @default.
• W3036404411 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/32569745" @default.
• W3036404411 hasPublicationYear "2020" @default.
• W3036404411 type Work @default.
• W3036404411 sameAs 3036404411 @default.
• W3036404411 citedByCount "21" @default.
• W3036404411 countsByYear W30364044112020 @default.
• W3036404411 countsByYear W30364044112021 @default.
• W3036404411 countsByYear W30364044112022 @default.
• W3036404411 countsByYear W30364044112023 @default.
• W3036404411 crossrefType "journal-article" @default.
• W3036404411 hasAuthorship W3036404411A5015680529 @default.
• W3036404411 hasAuthorship W3036404411A5021167805 @default.
• W3036404411 hasAuthorship W3036404411A5026956570 @default.
• W3036404411 hasAuthorship W3036404411A5027346080 @default.
• W3036404411 hasAuthorship W3036404411A5030497987 @default.
• W3036404411 hasAuthorship W3036404411A5041427467 @default.
• W3036404411 hasAuthorship W3036404411A5058485400 @default.
• W3036404411 hasAuthorship W3036404411A5087060483 @default.
• W3036404411 hasAuthorship W3036404411A5091612311 @default.
• W3036404411 hasBestOaLocation W30364044112 @default.
• W3036404411 hasConcept C104317684 @default.
• W3036404411 hasConcept C10853874 @default.
• W3036404411 hasConcept C153957851 @default.
• W3036404411 hasConcept C2776165026 @default.
• W3036404411 hasConcept C2779315393 @default.
• W3036404411 hasConcept C515207424 @default.
• W3036404411 hasConcept C547475151 @default.
• W3036404411 hasConcept C55493867 @default.
• W3036404411 hasConcept C67705224 @default.
• W3036404411 hasConcept C70721500 @default.
• W3036404411 hasConcept C86803240 @default.
• W3036404411 hasConceptScore W3036404411C104317684 @default.
• W3036404411 hasConceptScore W3036404411C10853874 @default.
• W3036404411 hasConceptScore W3036404411C153957851 @default.
• W3036404411 hasConceptScore W3036404411C2776165026 @default.
• W3036404411 hasConceptScore W3036404411C2779315393 @default.
• W3036404411 hasConceptScore W3036404411C515207424 @default.
• W3036404411 hasConceptScore W3036404411C547475151 @default.
• W3036404411 hasConceptScore W3036404411C55493867 @default.
• W3036404411 hasConceptScore W3036404411C67705224 @default.
• W3036404411 hasConceptScore W3036404411C70721500 @default.
• W3036404411 hasConceptScore W3036404411C86803240 @default.
• W3036404411 hasFunder F4320306915 @default.

• next