Fig. 5: Secondary structure analysis of PARP1 upon NAD+ addition.
From: Real-time monitoring of PARP1-dependent PARylation by ATR-FTIR spectroscopy
a Comparison of PARP1 bound to immobilised DNAblunt, DNA3’P, DNA5’P or DNAnick. Difference spectra of amide I bands of PARP1 before and after the addition of 100 µM NAD+ (0, 1 and 2 min) were calculated. Average curves of n = 3 independent experiments are plotted, respectively. SDs are shown in Supplementary Fig. 4a. Positive difference bands distinct from PAR bands are indicated in red. b Spectra of unlabelled and isotopically labelled 13C,15N-ATP. c Strategy for the enzymatic synthesis of isotopically labelled 13C,15N-NAD+. d Difference spectra of amide I bands of PARP1 bound to DNAblunt before and after the addition of 100 µM 13C,15N-NAD+ (0, 1 and 2 min) were calculated. Average curves of n = 3 independent experiments are plotted. SDs are shown in Supplementary Fig. 4d. The positive difference band distinct from the PAR band is indicated in red. Source data are provided as a Source Data file.
