Lanthanide fluorescence studies of transfer RNAf(met) conformation

The possible difference in conformation between aminoacylated and deacylated tRNA is examined using the optical and photochemical properties of the 4-thiouridine residue of E coli tRNAf(Met). No differences were seen between fMet-tRNAf(Met) and tRNAf(Met) observing the native fluorescence of 4-thiouridine, energy transfer from 4-thiouridine to the bound lanthanide ions, Tb3+ or Eu3+, or the rates of the photochemical cross-linking reaction of 4-thiourdine. While these results do not necessarily mean that there is no conformational difference between the aminoacylated and deacylated species, they do restrict the possible nature and magnitude of any conformational difference between the two species. In addition, preliminary thermal denaturation studies of tRNAf(Met), monitoring 4-thiouridine emission and energy transfer to Tb3+, indicate an unexplained melting phenomenon near 25 degrees C in the presence of Mg2+.