Fig. 4: 1H-PHIP effect observed for (C14a)Hpro-R.
a–d, The 1H spectra acquired for identification of the (C14a)Hpro-R PHIP effect, using four different experiments: 1H-experiments without 13C filtration (a and b; sequence in Supplementary Fig. 1) and with 13C filtration (c and d; sequence in Supplementary Fig. 3) were acquired with (a and c) or without (b and d) 13C decoupling (dec.) during acquisition. Overlays of experiments using p-H2 (magenta, 99% enrichment) or n-H2 (black) are shown in all cases, and the signal region for (C14a)Hpro-R in the free form of 13CH2=H4MPT (not bound to Hmd) is highlighted by green traces and amplified in the inserts. In all cases, high concentration samples were used (sample conditions stated in Fig. 3 legend), experiments were performed at 327 K and signals were accumulated over 16 scans (four treatments with p-H2 or n-H2, with an accumulation of four scans after each treatment). Formaldehyde-13C (13CH2=O) was added to improve sample stability. Different signal intensities for formaldehyde result from different degrees of formaldehyde deuteration due to the isotope exchange promoted by Hmd.
