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Determination of and Coupling Constants in 13C-Labeled Nucleic Acids Using Constant-Time HMQC

✍ Scribed by Weidong Hu; Serge Bouaziz; Eugene Skripkin; Abdelali Kettani

Publisher
Elsevier Science
Year
1999
Tongue
English
Weight
83 KB
Volume
139
Category
Article
ISSN
1090-7807

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✦ Synopsis

A novel 1 H-13 C correlated two-dimensional experiment, CT-HMQC-J, for the measurement of three-bond proton-phosphorus coupling constants in 13 C-labeled DNA is described. The experiment is based on the intensity difference of 1 H-13 C cross peaks in the presence and absence of the proton-phosphorus coupling interaction during the constant-time period in HMQC experiment. The 3 J(H, P) coupling constants can be easily extracted from the intensity ratios of the two experiments. The method has been applied to a uniformly 13 C, 15 N-labeled d(GGAGGAT) 7-mer DNA sample. The proton-phosphorus coupling constants determined from CT-HMQC-J, together with the other three-bond coupling constants, are used to determine ␤ and ⑀ torsion angles. The introduction of ␤ and ⑀ restraints has improved the convergence as well as the quality of d(GGAGGAT) structure.

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