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The 13C resonance frequency is 75.46 MHz and the data was collected at 273 K using a spectral width of 18.8 kHz, 1130 complex data points, acquisition time 30 ms, contact time 2 ms. A total of 1024 scans were acquired with a relaxation delay 4 s.
Typically 128 free induction decays (FIDs) were collected into 32k data points using a spectral width of 12 ppm.
For each acquisition, 128 transient free induction decays (FID) were collected into 32,768 data points using a spectral width of 20 ppm.
All data were collected using a spectral width of 20.0 ppm.
One-dimensional C NMR spectra were obtained using a spectral width of 24 kHz with 64 K data points.
One-dimensional H spectra were acquired using a spectral width of 5000 Hz and 32 K data points.
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For F MRS, a simple pulse-acquire sequence was used with a spectral width of 10 kHz, a time domain of 4 K points, a rectangular radiofrequency pulse of length 20 μs (ca. 70° flip angle at the centre of the surface coil), and a repetition time TR=0.5 s.
The HSQC-TOCSY spectra of 1, 2 and 3 were measured using the sequence covering a spectral width of 2762 Hz in F2 and 6667, 6849 and 6667 Hz in F1 with 1024 data points using 32, 32 and 64 transients for each of the 512, 460 and 512 t1 increments.
J HNHα couplings were obtained using HNHA experiments (Vuister and Bax, 1993), which were performed on the free and bound states using a 700 MHz spectrometer and a spectral width of 1454 Hz with 68 points in N and 9800 Hz with 72 80 points in the indirect H dimension.
For the assignment of the free state at pH 5.0 and 37°C, we used a previously published assignment (Ohkubo et al., 1991; Hagan and et al., 2010), which was confirmed using HNCA measurements, which was run with a spectral width of 1561 Hz and 68 points in the N dimension, and a spectral width of 5456 Hz and 64 points in the C dimension (Grzesiek and Bax, 1992).
A 90° excitation pulse was used to record 256 FID's with a spectral width of 8389 Hz averaged into 32 k data points with an acquisition time of 3.91 s.
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using a spectral wave
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using a spectral technique
using a spectral conditioning
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