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MQ-hCN-based Pulse Sequences for the Measurement of 13C1'-1H1', 13C1'-15N, 1H1'-15N, 13C1'-13C2', 1H1'-13C2',13C6/8-1H6/8, 13C6/8-15N, 1H6/8-15N, 13C6-13C5, 1H6-13C5 Dipolar Couplings in 13C, 15N-labeled DNA (And RNA)

Jiangli Yan, Takeshi Corpora, Padmanava Pradhan, John H Bushweller

J Biomol NMR. 2002 Jan;22(1):9-20.

PMID: 11885985

Abstract:

A suite of multiple quantum (MQ) HCN-based pulse sequences has been developed for the purpose of collecting dipolar coupling data in labeled nucleic acids. All the pulse sequences are based on the robust MQ-HCN experiment which has been utilized for assignment purposes in labeled nucleic acids for a number of years and provides much-needed resolution for the dipolar coupling measurements. We have attempted to collect multiple couplings centered on the 13C1' and 13C6/8 positions. Six pulse sequences are described, one each for measurement of one-bond 13C1'-1H1' and 13C6/8-1H6/8 couplings, one for measurement of one-bond 13C1'-15N and two-bond 1H1'-15N couplings, one for measurement of one-bond 13C6/8-15N and two-bond 1H6/8-15N couplings, one for measurement of one-bond 13C1'- 13C2' and two-bond 1H1'-13C2' couplings, and one for measurement of one-bond 13C6-13C5 and two-bond 1H6-13C5 couplings in the bases of C and T. These sequences are demonstrated for a labeled 18 bp DNA duplex in a 47 kDa ternary complex of DNA, CBFbeta, and the CBFalpha Runt domain, thus clearly demonstrating the robustness of the pulse sequences even for a very large complex.

Chemicals Related in the Paper:

Catalog Number Product Name Structure CAS Number Price
AP202407305 L-Valine-13C5,15N L-Valine-13C5,15N 202407-30-5 Price
IAR42410616 Fmoc-Ile-OH-13C6,15N Fmoc-Ile-OH-13C6,15N Price
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