5 mm tci probe

¹H NMR and 2D ROESY NMR spectra were performed on a Bruker Advance III HD spectrometer (600 MHz, Bruker BioSpin, Switzerland) fitted with a 5-mm TCI probe at 298 K, and TMS used as the calibration signal. The solvent in the experiment was D₂O (99%), CDCl₃ (99%), or DMSO-d₆ (99%).

One-dimensional ¹H-NMR spectra were recorded with a Bruker Avance III 500-MHz NMR spectrometer at 298 K (Bruker, Rheinstetten, Germany). All other NMR experiments were carried out with a Bruker Avance III 700-MHz NMR spectrometer, equipped with a cryogenically cooled, 5-mm TCI probe at 298 K. Samples containing 20–40 mg·mL⁻¹ NQO1 or NQO1 P187S in 50 mm Hepes (pH 6.5) in 90% H₂O and 10% D₂O were used for NMR measurements. For ¹⁵N relaxation measurements, series of 10 interleaved, relaxation-edited, ¹H/¹⁵N-HSQC spectra were recorded. The spectra were processed with nmrpipe [34 (link)] and analyzed with ccpnmr [35 (link)]. Rotational correlation times were calculated from the ¹⁵N T₁/T₂ ratios as previously described [36 (link)].

A. fumigatus conidia were incubated at room temperature for 1 h with 500 uM NAD⁺ in NMR reaction buffer (25 mM sodium phosphate pH 5.8, 50 mM NaCl, 5 % (v/v) D2O) in a final volume of 500 µl. Subsequently the conidia were removed by centrifugation followed by filtration through a 10 kDa MWCO Amicon® Ultra Centrifugal filter (Merck Millipore). The reaction products where identified by NMR and data were collected on a Bruker Ascend 850 MHz instrument fitted with a cryogenically cooled triple resonance 5 mm TCI probe with pulse filed gradients along the z-axis at 23 °C. The reaction products were identified by ¹H NMR using the pulse sequence zgesgppe allowing water suppression using excitation sculpting with pulse field gradients and perfect echo. The spectra were acquired with 16 scans and a recovery delay of 3.9 seconds