Comparison of H5 and H8 relaxation rates of a 2H/13C/15N labeled RNA oligonucleotide with selective protonation at C5 and C8.

Uniformly 13C/15N enriched ribonucleotide monophosphates have been prepared with extensive deuterium enrichment of the non-exchangeable positions. The purine C8 and pyrimidine C5 base positions were selectively protonated prior to incorporation of the individual nucleotide triphosphates into an RNA oligonucleotide. The longitudinal and transverse relaxation rates of the H8 and H5 resonances of this deuterated molecule were compared with the relaxation rates of the corresponding protonated, 13C/15N enriched RNA molecule. Deuteration disrupts the efficiency of 1H-1H dipolar relaxation and reduces the longitudinal and transverse magnetization relaxation rates on average to 25% and 68%, respectively, of the values measured for the non-deuterated RNA molecule. Importantly, the longitudinal relaxation rates remain sufficiently rapid (> 1s[-1]) to permit the use of short recovery delays in multidimensional NMR experiments without significant loss of sensitivity.