JE-TROSY: combined J- and TROSY-spectroscopy for the measurement of one-bond couplings in macromolecules
B. Luy, John Marino
Since the recent introduction of the use of residual dipolar couplings in high-resolution NMR studies of macromolecules, there has been a renewed interest in rapid, sensitive methods for the measurement of 15N-1H} and 13C-1H} one-bond couplings. For large macromolecules and macromolecular complexes (M.W. > 25 kDa), methods for the determination of one-bond couplings that rely on the measurement of the displacement between the two components of an HX coupled cross peak, are often unreliable since differential relaxation significantly broadens one of the two multiplet components needed for accurate measurements. In this communication, a method to measure 15N-1H} and 13C-1H} one-bond couplings is presented that utilizes J-evolution of the sharpest multiplet component selected in a heteronuclear transverse relaxation optimized spectroscopy (TROSY) experiment. It is demonstrated that this method circumvents this problem of differential line-broadening and provides resolved, sensitive spectra from which J-couplings can be accurately measured in large macromolecular systems.
Journal of Magnetic Resonance Imaging
D-coupling, J-coupling, NMR, RNA, TROSY
and Marino, J.
JE-TROSY: combined J- and TROSY-spectroscopy for the measurement of one-bond couplings in macromolecules, Journal of Magnetic Resonance Imaging
(Accessed December 3, 2023)