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• W2046187851 abstract "Conformational transitions of Phe-tRNA(Phe) that take place during elongation factor Tu (EF-Tu)-dependent binding to the A site of Escherichia coli ribosomes were followed by transient fluorescence measurements. The fluorescence signal of proflavin replacing dihydrouracil at position 16 or 17 in yeast tRNA(Phe) was utilized to monitor changes of the conformation of the D loop. The ternary complex EF-Tu.GTP.Phe-TRNA(Phe)(Pf16/17) was purified by gel filtration. Upon binding of the complex to the A site of poly(U)-programmed, P-site-blocked ribosomes, the fluorescence changes in several steps. First, the rapid formation of an initial complex gives rise to a small fluorescence increase. Subsequent codon-anticodon recognition leads to a conformational rearrangement of the D loop of the tRNA that is reflected in a major fluorescence increase. Fluorescence-quenching data indicate an unfolding of the D loop in this state. The latter conformational state is short-lived, and the aminoacyl-tRNA refolds during the following rearrangement that occurs after GTP hydrolysis and accompanies the release of the aminoacyl-tRNA from EF-Tu.GDP and/or its accommodation in the A site. Further experiments show that the status of the P site influences the binding to the A site in that the two rearrangement steps are slowed down when the P site is unoccupied and even more so when it is occupied with the near-cognate tRNA(Leu2). In contrast, the occupancy of the E site has no influence on A-site binding, and vice versa, thus excluding any coupling between the two sites." @default.
• W2046187851 created "2016-06-24" @default.
• W2046187851 creator A5017069360 @default.
• W2046187851 creator A5024960912 @default.
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• W2046187851 date "1994-10-11" @default.
• W2046187851 modified "2023-09-25" @default.
• W2046187851 title "Transient Conformational States of Aminoacyl-tRNA during Ribosome Binding Catalyzed by Elongation Factor Tu" @default.
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• W2046187851 doi "https://doi.org/10.1021/bi00206a033" @default.
• W2046187851 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/7918447" @default.
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