Determination of Backbone Angle ψ in Proteins Using a TROSY-Based α/β-HN(CO)CA-J Experiment

Transverse relaxation-optimized NMR experiment (TROSY) for the measurement of three-bond scalar coupling constant between ¹H^α_i−1 and ¹⁵N_i defining the dihedral angle ψ is described. The triple-spin-state-selective experiment allows measurement of ³J_H^αN from ¹³C^α, ¹⁵N, and ¹H^N correlation spectra H₂O with minimum resonance overlap. Transverse relaxation of ¹³C^α spin is minimized by using spin-state-selective filtering and by acquiring a signal longer in ¹⁵N-dimension in a manner of semi-constant-time TROSY evolution. The ³J_H^αN values obtained with the proposed α/β-HN(CO)CA-J TROSY scheme are in good agreement with the values measured earlier from ubiquitin in D₂O using the HCACO[N] experiment.