A nuclear magnetic resonance study of secondary and tertiary structure in yeast tRNAPhe.

We present experimental evidence which confirms recently proposed ring current prediction methods for assigning hydrogen-bond proton nuclear magnetic resonance (NMR) spectra from tRNA (Robillard, G. T., Tarr, C. E., Vosman, F., & Berendsen, H. J. C. (1976) Nature (London) 262, 363-369; Robillard, G. T., Tarr, C. E., Vosman, F., & Sussman, J. L. (1977) Biophys. Chem. 6, 291-298). The evidence is a series of temperature-dependent studies on yeast tRNAPhe monitoring both the high- and low-field NMR spectral regions, which are correlated with independent optical and temperature-jump (temp-jump) studies performed under identical ionic strength conditions. Using assignments derived from the new prediction methods, the melting patterns of the hydrogen-bonded resonances agree with those expected on the basis of optical, temp-jump, and NMR studies on the high-field spectral region. The implication of these results is that previous assignment procedures are at least partially incorrect and, therefore, studies based on those procedures must be reexamined.