Separation of formyl-methionyl transfer RNA, methionyl transfer RNA, and transfer RNAfmet using mixed-mode high-performance liquid chromatography on C6-modified aminopropylsilyl-hypersil
✍ Scribed by Robert D. Ricker; Akira Kaji
- Publisher
- Elsevier Science
- Year
- 1988
- Tongue
- English
- Weight
- 672 KB
- Volume
- 175
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
Preparative amounts of formyl-methionyl-tRNAfmet, methionyl-tRNAfma, and tRNAfma were separated from each other with baseline resolution in 30 min using mixed-mode HPLC on hexanoic anhydride-modified aminopropylsilyl-Hypemil2.
Pure tRNAfmti was aminoacylated with [3'S]methionine in the presence or absence of a formyl donor and was immediately fractionated on the column. Two isoacceptors, tRNAima and tRNAima, as well as aminoacyl-tRNA synthetases were clearly separated from each other. The purified f135S]-methionyl-tRNA was biologically active in that as much as 98% could be bound to ribosomes in response to AUGUAA in vitro. Formyl-methionine was released from this complex by the action of termination factor and >92% of bound formyl-methionine was released by puromycin. 0 1988 Academic presg ITIC.