Figure 8. ³¹P-NMR spectra of RsBluE reaction products.

Representative ³¹P-NMR spectra of duplicate independent experiments. Reaction mixtures containing MgCl₂ (1 mM), ATP (3 mM), L-Thr (6 mM), and 10 μL of detergent-free RsBluE protein (11 μM) were incubated at 25°C for 1 h. A. No enzyme reactions with AMP, sodium ortho-phosphate (Pi), sodium pyrophosphate (PPi), and sodium polytriphosphate (PPPi) standards. B. No enzyme reactions with L-Thr-P and ATP standards. C. Reaction containing ATP and RsBluE. D. Reaction containing ATP, L-Thr, and RsBluE. E. Reaction containing ATP, L-Thr, and extracts from cells carrying pTEV5 empty vector. Conditions used for the acquisition of the spectra are described in the Materials and Methods section.