Reversed-Phase Chromatography Systems for Transfer Ribonucleic Acids — Preparatory-Scale Methods

Kelmers, A.D.; Weeren, H.O.; Weiß, J.; Pearson, R.L.; Stulberg, M.P.; Novelli, G. David

doi:10.1016/b978-0-12-504180-5.50007-9

Cited by 3 publications

(4 citation statements)

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“…coli aminoacyl-tRNA synthases obtained by the fractionation of a cell lysate with ammonium sulfate, which was reported to contain practically all the aminoacyl-tRNA synthases [5]. The enzymatic activity of this preparation was determined by its ability to aminoacylate total E .…”

Section: Resultsmentioning

confidence: 99%

“…The following preparations were used in this work: A total fraction of aminoacyl-tRNA synthases from E. coli was isolated by the method [5], and its enzymatic activity was determined from the ability to attach [ 3 H]leucine and [ 35 S]methionine to total E. coli tRNA as described in [6]. The starting nucleosides were provided by the AZT Association company (Moscow).…”

Section: Methodsmentioning

confidence: 99%

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Enzymatic Synthesis of Bis(5′-Nucleosidyl) Tetra- and Triphosphates

et al. 2005

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The total fraction of aminoacyl-tRNA synthases from Escherichia coli has been shown to catalyze the synthesis of the bis(5'-nucleosidyl) oligophosphates Ap4AZT, Ap4d4T, Ap43TC, and Ap4ACV, as well as Ap3AZT and Ap3d4T, from [alpha-32P]ATP and the corresponding nucleoside-5'-tri(or di)phosphate. The resulting compounds, characterized by HPLC, are resistant to alkaline phosphatase. Ap4AZT, Ap4d4T, and Ap43TC are formed with approximately equal efficiency, whereas the efficiencies of the synthesis of Ap4ACV, Ap3AZT, and Ap3d4T are three- to fivefold lower. The English version of the paper: Russian Journal of Bioorganic Chemistry, 2005, vol. 31, no. 6; see also http://www.maik.ru.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Enzymatic Synthesis of Bis(5′-Nucleosidyl) Tetra- and Triphosphates

et al. 2005

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“…Purified aminoacyl-tRNA synthetases from E.coli were a gift of C.Dingwell. An E.coli extract containing all aminoacyl-tRNA synthetases was prepared according to Kelmers et al (13). E.coli tRNA (type XXI from Sigma) was deacylated by incubation at 370C for 1 hour in 1 M Tris, pH 8.8, and extensively dialyzed before use.…”

Section: Methodsmentioning

confidence: 99%

Selective³²P-labelling of individual species in a total tRNA population

Traboni

Cortese²,

Salvatore

1980

Nucl Acids Res

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A simple procedure to label individual tRNA species in a total tRNA preparation has been developed. The principle of the method is as follows: total crude tRNA (from E. coli) is incubated in the presence of a crude aminoacyl-tRNA synthetase preparation, containing most aminoacyl-tRNA synthetases and only one specific amino acid corresponding to the tRNA species which is intended to be labelled. This achieves the purpose of charging the desired tRNA species thereby protecting its 3'OH-terminus; obviously all the other tRNA species will have a free 3'OH group. Periodate oxidation, followed by beta-elimination, destroys any free 3'OH. After deacylation of the specific aminoacylated tRNA at pH 8.8 the only free 3'OH group will be the one of the desired tRNA species. High specific activity (32P)-pCp is ligated to this 3'OH by means of T4-RNA ligase. Two-dimensional polyacrylamide gel electrophoresis (2D-PGE) and sequence analysis of the isolated tRNA show that the method is very specific. Individually labelled tRNA species can be used as probes for cloning tRNA genes.

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“…These include gel filtration, affinity and ion exchange [1][2][3][4][5][6][7][8][9][10][11][12][13]. These include gel filtration, affinity and ion exchange [1][2][3][4][5][6][7][8][9][10][11][12][13].…”

Section: Development Of Dna and Rna Chromatographymentioning

confidence: 99%