Fractionation of rat liver tRNA by reversed-phase high performance liquid chromatography: isolation of Iso-tRNAs(Pro).
This paper illustrates the fractionation of cytoplasmic transfer ribonucleic acid from rat liver by reversed-phase high performance liquid chromatography using a gradient of acetonitrile/ammonium acetate. The procedure is fast, highly reproducible, and gives an excellent resolution of the numerous tRNA population: about 50 peaks with area peak percentages ranging from 0.001 to 5 can be monitored. Uncharged tRNA preparations exhibited a chromatographic profile different from aminoacylated tRNA, thus suggesting a possible strategy to distinguish between aminoacylated and nonacylated tRNA species. Moreover, a first approach to map the HPLC peaks was attempted by chromatographing preparations of tRNA which had been aminoacylated with individual 3H-labeled aminoacids. Here is reported the case of tRNA(Pro), which gave three well separated radioactive peaks, most likely corresponding to tRNA(Pro) isoacceptor species.
Kanduc
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Dept. of Biochemistry and Molecular Biology, Faculty of Sciences, University of Bari.
D000085:Acetates; D000097:Acetonitriles; D000215:Acylation; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008099:Liver / Q000737:chemistry*; D012361:RNA, Transfer, Pro / Q000302:isolation & purification*; D051381:Rats; D015203:Reproducibility of Results
D000085:Acetates; D000097:Acetonitriles; D012361:RNA, Transfer, Pro; C018824:ammonium acetate; C032159:acetonitrile