Improvements for measuring 1H-1H coupling constants in DNA via new stripe-COSY and superstripe-COSY pulse sequences combined with a novel strategy of selective deuteration

Journal of Magnetic Resonance
J YangM A Kennedy

Abstract

Three bond proton-proton vicinal coupling constants are of potential value for analyzing sugar conformations in DNA. However, self-cancellation in antiphase cross peaks and modulation of peak splittings by transverse cross relaxation can alter the apparent coupling constants such that they do not accurately reflect the sugar conformations. Transverse cross relaxation is most effective between strongly coupled geminal proton pairs. Here we report the use of stereospecific deuteration at the H2" position in the A5 and A6 residues in the 12 base pair DNA sequence [d(CGCGAATTCGCG)2] as a means of investigating the effect of transverse cross relaxation on P.E.COSY type cross peaks. Deuteration of the H2" proton is expected to reduce the transverse cross relaxation rate by the square of ratio of the proton to deuteron gyromagnetic ratios, i.e., by a factor of 42. Additionally, a striking eight- to ninefold increase in the signal intensity was observed for cross peaks involving the remaining H2' proton resulting from diminished dipolar relaxation. Further improvements in signal-to-noise ratio were realized by collecting P.E.COSY spectra in strips, using an experiment referred to as stripe-COSY, employing selective excitation pulses wh...Continue Reading

Citations

Sep 22, 1999·European Journal of Biochemistry·R P OjhaR H Sarma
Feb 24, 2001·Nucleosides, Nucleotides & Nucleic Acids·A FöldesiJ Chattopadhyaya
Apr 6, 2005·Journal of Magnetic Resonance·Scott A BradleyChad E Hadden
Jul 1, 2020·Journal of Biomolecular Structure & Dynamics·Mariana de Oliveira Tonelli NogueiraJosé Daniel Figueroa-Villar
Jul 14, 2020·Journal of Biomolecular Structure & Dynamics·Priscila Ivo Rubim de SantanaJosé Daniel Figueroa-Villar

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