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
Date Published2003 Jul
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

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