(a) 2D MEIOSIS pulse sequence for simultaneous acquisition of four experiments: DARR, N(CA)CX, NCO, and CA(N)CO. (b) MEIOSIS pulse sequence with CON and CAN transfers inverted. The phase cycle of 2D MEIOSIS and conventional pulse sequences of Fig. 1S of the supplementary material 36 are given by: ϕ1 = (y)4, (−y)4, ϕ2 = (x, x, −x, −x)2, ϕ3 = (y, −y)4, ϕ4 = (y, y, −y, −y)2, and ϕrec = (x, −x, −x, x, −x, x, x, −x). For indirect dimension, states mode acquisition is obtained by alternating the ϕ* between x and y.
(a) Experimental plot of ubiquitin showing the normalized intensities of four pathways during NCA (or CAN) transfer, as a function of specific-CP contact time (τ). Each data point is obtained by integrated intensities of the 1D spectra shown in (b), by using the pulse sequence of Fig. 1(a) with , , and DARR mixing period set to zero. The maximum intensity of each pathway is normalized to one. The data points (a) and the corresponding spectra in (b) are color-coded according to respective pathways shown in Fig. 1 . Open and closed orange colored circles indicate Cα (50–75 ppm) and aliphatic carbon region (0–75 ppm) intensities. The arrow indicates the optimal contact time (2916 μs) for maximum NCA and CAN transfer. The residual magnetization at 2916 μs corresponds to ∼34% for 15N and ∼43% for the 13Cα (50–75 ppm) and 54% for 13C aliphatic (0–75 ppm) resonances. The residual magnetization of C′ region is less than 5% due to insufficient spin lock field.
Simultaneous acquisition of four 2D spectra using the MEIOSIS pulse sequence reported in Fig. 1(a) .
Three-dimensional MEIOSIS pulse sequence for simultaneous acquisition of three 3D spectra: NCACX, NCOCX, and CAN(CO)CX. The phase cycle is given by ϕ1 = (y, −y)2, ϕ2 = x, x, −x, −x, and ϕrec = x, −x, −x, x. Sates mode acquisition of indirect dimensions is obtained by alternating ϕ2, and ϕ* phases between x and y.
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