Europium as a fluorescent probe of transfer RNA structure

The binding of europium(III) to Escherichia coli tRNA-fMet,Glu and to unfractionated E. coli tRNA has been investigated by using the 4-thiouridine sensitization of europium 5-Do yields 7-F1 emission and changes in the lifetime of the 5-Do state of europium reported earlier (J. M. Wolfson and D. R. Kearns (1974), J. Am. Chem. Soc. 96, 3653). Binding of the first 3-4 europium ions is independent and sequential, approximately 600 times stronger than the magnesium binding, and the binding sites are located near the 4-thiouridine residue found at position 8 in a number of E. coli tRNA. Competition experiments suggest the strong binding sites are the same for magnesium and europium. The europium binding properties of both unfractionated E. coli tRNA and purified tRNA-fMet are quite similar, indicating that the location of the strong binding sites and their binding constants are nearly the same for a large group of tRNA. The europium binding properties of native and denatured tRNA are quite different, however.