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

Barry I. Schweitzer, Kevin H. Gardner, Gregory Tucker-Kellogg

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)180-188
Number of pages9
JournalJournal of biomolecular NMR
Volume6
Issue number2
DOIs
StatePublished - Sep 1995

Keywords

  • Cadmium
  • Cytosine arabinoside
  • DNA
  • Hetero TOCSY
  • Heteronuclear
  • LAC9
  • Phosphorus
  • Relaxation

ASJC Scopus subject areas

  • Biochemistry
  • Spectroscopy

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