Drx 700 mhz spectrometer

NMR spectra were acquired at 298 K on a Bruker DRX 700 MHz spectrometer operating at the proton frequencies of 700.13 MHz (16.44 T) on uniformly ¹⁵N- enriched samples of ca. 0.5 mM UreG in 20 mM TrisHCl buffer at pH 8, containing 150 mM NaCl, 1 mM TCEP, in the absence and in the presence of 2 mM SDS. The spectrometer was equipped with a triple-resonance 5-mm TCI cryo-probe with pulse field gradients along the z-axis. The spectra were acquired using 2048 × 128 number of complex points and the hsqcfpf3gpphwg Bruker library pulse sequence.

NMR samples for C-Ps standards were prepared by dissolving C-Ps1 (SSI, CWPS), or a mixture of C-Ps1 and C-Ps2 (SSI, CWPS-Multi) in the Deuterium oxide (D₂O, 99.96% atom D) containing 0.01% (v/v) dimethylsulfoxide (DMSO), used as an internal intensity standard and 0.01% (w/v) DSS-d₆, used as an internal chemical-shift standard. The concentration of the C-Ps standard in the NMR sample is about 5 mg/mL. ¹H NMR spectra were acquired at 49 °C on a Varian DD2 600 MHz spectrometer with a 5 mm HCN CryoProbe and by running the VnmrJ version 4.2. ¹H NMR spectra were recorded using a standard one-pulse experiment with 32,768 data points, a spectral width of 10 ppm, and 4 scans. For the quantitative study, relaxation time was set to 57 s to ensure a full recovery of each signal. One-dimensional ³¹P NMR spectra were acquired at 25 °C on the Bruker DRX 700 MHz spectrometer with an HCP Cryoprobe. Suitable signal-to-noise ³¹P spectra were recorded, accumulating approximately 256 scans with 32,768 data points and a spectral width of 75 ppm, weighted with 2.0 Hz line broadening, and were Fourier transformed. The ratio of C-Ps1 and C-Ps2 in the CWPS-Multi was provided by SSI and confirmed by ³¹P NMR (Supplementary Figure S2). Quantitative NMR analysis for C-Ps was performed based on the resonance integration of choline signals at 3.22 ppm and the DMSO signal at 2.71 ppm (Supplementary Figure S3).