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.
Dept. of Biochemistry and Molecular Biology, Faculty of Sciences, University of Bari.
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Kanduc
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