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Journal of Biomolecular NMR

, Volume 6, Issue 2, pp 180–188 | Cite as

HeteroTOCSY-based experiments for measuring heteronuclear relaxation in nucleic acids and proteins

  • Barry I. Schweitzer
  • Kevin H. Gardner
  • Gregory Tucker-Kellogg
Research Paper

Summary

While both 31P and 113Cd are present at locations of interest in many different macromolecular systems, heteronuclear-detected relaxation measurements on these nuclei have been restrained by limitations in either resolution or signal-to-noise ratio. We have developed hetero TOCSY-based methods to overcome both of these problems. Two-dimensional versions of these experiments were utilized to measure 31P T1 and T2 values in DNA oligonucleotides; the additional resolution offered by a second dimension allowed determination of these values for most of the 31P resonances in a DNA dodecamer. The results from the experiments indicated that there was little significant variation in T1 values for the different phosphates in the DNA dodecamer; however, the T2 values showed a clear pattern, with lower values in the interior of the sequence than at the ends of the helix. Furthermore, a significant correlation between 31P chemical shifts and T2 values was observed. One-dimensional, frequency-selective versions of these experiments were also developed for use on systems containing a smaller number of heteronuclear spins. These methods were applied to investigate the heteronuclear relaxation properties of 113Cd in 113Cd2LAC9(61), a Cys6Zn2 DNA-binding domain. Data from the experiments confirm biochemical evidence that more significant differences occur in the metal-protein interactions between the two metal-binding sites than has been previously identified for proteins containing this motif.

Keywords

DNA Phosphorus Cadmium Heteronuclear Relaxation Hetero TOCSY Cytosine arabinoside LAC9 

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Copyright information

© ESCOM Science Publishers B.V 1995

Authors and Affiliations

  • Barry I. Schweitzer
    • 1
  • Kevin H. Gardner
    • 3
  • Gregory Tucker-Kellogg
    • 2
    • 3
  1. 1.Department of PediatricsYale University School of MedicineNew HavenU.S.A.
  2. 2.Department of ChemistryYale UniversityNew HavenU.S.A.
  3. 3.Department of Molecular Biophysics and BiochemistryYale UniversityNew HavenU.S.A.

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