Fig. 3: Experimental setup for in-cyclo NMR with ATP regeneration and phosphate removal system.
From: Mechanism of ATP hydrolysis in the Hsp70 BiP nucleotide-binding domain
a Reaction scheme of the ATP regeneration system (E1; pyruvate kinase) and phosphate removal system (E2; sucrose phosphorylase). b Series of 1D 1H NMR spectra of the ATP indole H8, recorded at an interval of 6 min. c Quantification of the ATP concentration from the 1D 1H NMR spectra shown in (b): Red crosses represent three independent experiments. d Same as (b) for pyruvate (left), PEP (middle) and glucose-1-phosphate (right). e Quantification of the pyruvate (black circle) and glucose-1-phosphate (lavender triangle) concentration from the 1D 1H NMR spectra shown in (d): the symbols represent three independent experiments. The black line shows a linear fit to determine the ATP consumption rate. f Molecular hydrolysis rate calculated from the in-cyclo data presented in (e). Data points represent three independent experiments. The bar represents mean and standard deviation.
