Sensitivity-enhanced double-TROSY experiment for simultaneous measurement of one-bond 15 N- 1 H, 15 N- 13 C and two-bond 1 H- 13 C couplings

Abstract

A recently published experiment for the measurement of 1HN, 1NC', and 2JHC' coupling constants [J. Am. Chem. Soc. 125 (2003) 11504] was modified to yield a double-TROSY experiment which selects 1 of the 16 multiplet components from a 15N-HSQC spectrum recorded of a uniformly 15N/ 13C-labelled protein. Subspectra containing any 1 of the 16 multiplet components can be generated allowing accurate coupling constant measurements. The experiment is sensitivity enhanced, turning all magnetization components precessing during the evolution time into observable magnetization during the detection time. The experiment is discussed with regard to the previously published α/β-filtered HN(α/β-NC'-J) experiment [J. Magn. Reson. 140 (1999), 32] which measures the same coupling constants.