Poster abstracts
Poster number 80 submitted by Jennifer Kist
Offline HPLC fractionation and LC-MS/MS characterization of tRNAs
Jennifer A. Kist (Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati), Cassandra Herbert (Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati), Scott Abernathy (Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati), Patrick A. Limbach (Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati)
Abstract:
Transfer ribonucleic acids (tRNAs) are adapter molecules acting as a carrier of amino acids to the growing polypeptide chain. Among all RNAs, tRNAs exhibit the largest number and the widest variety of modifications to ensure proper structure and function. Mass spectrometry methods are becoming increasingly important for the characterization of post-transcriptional modifications on ribonucleic acids. RNA Modification Mapping by LC-MS/MS is applicable to mixtures of RNAs (in particular, tRNAs) from bacterial and archaeal organisms. However, many organisms, especially eukaryotes, can transcribe hundreds or more individual tRNA sequences within the cell. Such mixtures are inherently difficult to accurately map to particular sequences given current technologies. In this work, a reverse phase (RP) HPLC method for fractionating tRNAs compatible with RNA modification mapping was developed. Yeast tRNA as well as tRNA from WM793 primary melanoma cells were both successfully fractionated into 4 fractions. The fractions were collected, washed, and ran on LC-MS for nucleoside analysis.
Keywords: tRNA, LC-MSMS, RP-HPLC