Gw. Kellogg et Bi. Schweitzer, 2-DIMENSIONAL AND 3-DIMENSIONAL P-31-DRIVEN NMR PROCEDURES FOR COMPLETE ASSIGNMENT OF BACKBONE RESONANCES IN OLIGODEOXYRIBONUCLEOTIDES, Journal of biomolecular NMR, 3(5), 1993, pp. 577-595
We describe a strategy for sequential assignment of P-31 and deoxyribo
se H-1 NMR resonances in oligode-oxyribonucleotides. The approach is b
ased on P-31-H-1 J-cross-polarization (heteroTOCSY) experiments, recen
tly demonstrated for the assignment of resonances in RNA [Kellogg, G.W
. (1992) J. Magn. Reson., 98, 176; Kellogg, G.W. et al. (1992) J. Am.
Chem. Soc., 114, 27271. Two-dimensional heteroTOCSY and heteroTOCSY-NO
ESY experiments are used to connect proton spin systems from adjacent
nucleotides in the dodecamer d(CGCGAATTCGCG)2 entirely on the basis of
through-bond scalar connectivities. All phosphorus resonances of the
dodecamer are assigned by this method, and many deoxyribose H-1 resona
nces can be assigned as well. A new three-dimensional heteroTOCSY-NOES
Y experiment is used for backbone proton 4', 5' and 5'' resonance assi
gnments, completing assignments begun on this molecule in 1983 [Hare,
D.R. et al. (1983) J. Mol Biol., 171, 3191. Numerical simulations of t
he time dependence of coherence transfer aid in the interpretation of
heteroTOCSY spectra of oligonucleotides and address the dependence of
heteroTOCSY and related spectra on structural features of nucleic acid
s. The possibility of a generalized backbone-driven H-1 and P-31 reson
ance-assignment strategy for oligonucleotides is discussed.