Nitrogen-15-labeled yeast tRNAPhe: double and two-dimensional heteronuclear NMR of guanosine and uracil ring NH groups

Abstract

5N1-Labeled hypoxanthine and 1,3-15N-labeled uracil were synthesized chemically and used to prepare labeled yeast tRNAPhe biosynthetically. Maps (500 MHz) of 15N chemical shift vs. proton chemical shift were obtained, for each ring NH group, by means of INDOR (difference heterodecoupling) and also by means of a proton-observe two-dimensional method involving coherences of forbidden resonances of the NH system. Resonances of GC11, T54-m1A58, GU4, and A psi 31 were confirmed, assigned, or reassigned. psi 39 was found to be in anti conformation, not syn as previously stated. Almost all the uracil NH group resonances could be separated, but most of the GC resonances are too close even in two dimensions to be separately resolved with the observed 20-Hz 15N line width.