Complete purification of tRNA, charged or modified with hydrophobic groups, by reversed-phase high-performance liquid chromatography on a C4/C18 column system
1994; Elsevier BV; Volume: 679; Issue: 1 Linguagem: Inglês
10.1016/0021-9673(94)80314-5
ISSN1873-3778
AutoresJ.R. Mesters, Erik L.H. Vorstenbosch, Aldo J. de Boer, Barend Kraal,
Tópico(s)Mass Spectrometry Techniques and Applications
ResumoPhe-tRNAPhe, N-acetyl-Phe-tRNAPhe and Leu-tRNALeu-4 (from brewer's yeast and Escherichia coli) were each separated with baseline resolution from the uncharged tRNA species by using a wide-pore C4 column and inverse salt gradient elution. The alterations at the 3′ end of the tRNAs result in a considerable shift of retention time on this column. The method is useful not only for obtaining tRNA preparations as required for poly(U) translational studies, but also for producing 20–50-mg amounts of tRNA for NMR and X-ray analysis. These aminoacylated species (charged by crude synthetase mixtures) can be purified from the crude tRNA mixtures in a one-step procedure.
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