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@article{314211,
author = {Fiala, Radovan and Czernek, Jiří and Sklenář, Vladimír},
article_location = {Nizozemí},
article_number = {4},
keywords = {C-13 CSA; C-13 relaxation; HCN experiments; RNA; TROSY},
language = {eng},
issn = {0925-2738},
journal = {Journal of Biomolecular NMR},
title = {Transverse relaxation optimized triple-resonance NMR experiments for nucleic acids},
volume = {16},
year = {2000}
}
TY - JOUR
ID - 314211
AU - Fiala, Radovan - Czernek, Jiří - Sklenář, Vladimír
PY - 2000
TI - Transverse relaxation optimized triple-resonance NMR experiments for nucleic acids
JF - Journal of Biomolecular NMR
VL - 16
IS - 4
SP - 291
EP - 291
PB - Kluwer/Escom
SN - 09252738
KW - C-13 CSA
KW - C-13 relaxation
KW - HCN experiments
KW - RNA
KW - TROSY
N2 - Triple resonance HCN and HCNCH experiments are reliable methods of establishing sugar-to-base connectivity in the NMR spectra of isotopicaly labeled oligonucleotides. However, with larger molecules the sensitivity of the experiments is drastically reduced due to relaxation processes. Since the polarization transfer between 13C and 15N nuclei relies on rather small heteronuclear coupling constants (11-12 Hz), the long evolution periods (up to 30-40 ms) in the pulse sequences cannot be avoided. Therefore any effort to enhance sensitivity has to concentrate on manipulating the spin system in such a way that the spin-spin relaxation rates would be minimized. In the present paper we analyze the efficiency of the two known approaches of relaxation rate control, namely the use of multiple-quantum coherence - MQ (Griffey and Redfield, 1987; Bax et al., 1989; Grzesiek and Bax, 1995) and of the relaxation interference between chemical shift anisotropy and dipolar relaxation - TROSY (Pervushin et al. 1997; Pervushin et al. 1998a; Pervushin et al. 1998b; Andresson et al. 1998; Rance et al. 1999; Brutcher et al. 1998; Salzman et al. 1998; Salzman et al. 1999). Both theoretical calculations and experimental results suggest that for the sugar moiety (H1'-C1'-N1/9) the MQ approach is clearly preferable. For the base moiety (H6/8-C6/8-N1/9) however, the TROSY shows results superior to the MQ suppression of the dipole-dipole relaxation at moderate magnetic fields (500 MHz) and the sensitivity improvement becomes dramatically more pronounced at very high fields (800 MHz). The pulse schemes of the triple-resonance HCN experiments with sensitivity optimized performance for unambiguous assignments of intra-residual sugar-to-base connectivities combining both approaches are presented.
ER -
FIALA, Radovan, Jiří CZERNEK a Vladimír SKLENÁŘ. Transverse relaxation optimized triple-resonance NMR experiments for nucleic acids. \textit{Journal of Biomolecular NMR}. Nizozemí: Kluwer/Escom, 2000, roč.~16, č.~4, s.~291-302. ISSN~0925-2738.
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