B. Brutscher et al., Base-type-selective high-resolution C-13 edited NOESY for sequential assignment of large RNAs, J BIOM NMR, 19(2), 2001, pp. 141-151
Extensive spectral overlap presents a major problem for the NMR study of la
rge RNAs. Here we present NMR techniques for resolution enhancement and spe
ctral simplification of fully C-13 labelled RNA. High-resolution H-1-C-13 c
orrelation spectra are obtained by combining TROSY-type experiments with mu
ltiple-band-selective homonuclear C-13 decoupling. An additional C-C filter
sequence performs base-type-selective spectral editing. Signal loss during
the filter is significantly reduced because of TROSY-type spin evolution.
These tools can be inserted in any C-13-edited multidimensional NMR experim
ent. As an example we have chosen the C-13-edited NOESY which is a crucial
experiment for sequential resonance assignment of RNA. Application to a 33-
nucleotide RNA aptamer and a 76-nucleotide tRNA illustrates the potential o
f this new methodology.