JE-TROSY: combined J- and TROSY-spectroscopy for the measurement of one-bond couplings in macromolecules.

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TitleJE-TROSY: combined J- and TROSY-spectroscopy for the measurement of one-bond couplings in macromolecules.
Publication TypeJournal Article
Year of Publication2003
AuthorsLuy, B, Marino, JP
JournalJ Magn Reson
Volume163
Issue1
Pagination92-8
Date Published2003 Jul
ISSN1090-7807
KeywordsAlgorithms, Bacterial Proteins, Biopolymers, Carbon Isotopes, Hydrogen Bonding, Macromolecular Substances, Nitrogen Isotopes, Nuclear Magnetic Resonance, Biomolecular, Oligonucleotides, Protein Binding, Protons, Reproducibility of Results, RNA, RNA-Binding Proteins, Sensitivity and Specificity, Signal Processing, Computer-Assisted, Spin Labels
Abstract

With the application of RDCs in high-resolution NMR studies of macromolecules, there has been an interest in the development of accurate, sensitive methods for measuring 15N-1H and 13C-1H one-bond coupling constants. Most methods for determining these couplings are based on the measurement of the displacement between cross-peak components in J-coupled spectra. However, for large macromolecules and macromolecular complexes, these methods are often unreliable since differential relaxation can significantly broaden one of the multiplet components (i.e., the anti-TROSY component) and thereby make accurate determination of its position difficult. To overcome this problem, a J-evolved transverse relaxation optimized (JE-TROSY) method is presented for the determination of one-bond couplings that involves J-evolution of the sharpest cross-peak multiplet component selected in a TROSY experiment. Couplings are measured from the displacement of the TROSY component in the additional J-evolution dimension relative to a zero frequency origin. The JE-TROSY method is demonstrated on uniformly labeled 15N, 13C-labeled RNA and peptide samples, as well as with an RNA-protein complex, in which the protein is uniformly 15N, 13C-labeled. In all cases, resolved, sensitive spectra are obtained from which heteronuclear one-bond J-couplings could be accurately and easily measured.

Alternate JournalJ. Magn. Reson.
PubMed ID12852911
Grant ListGM59107 / GM / NIGMS NIH HHS / United States