A single evolutionarily divergent mutation determines the different FAD‐binding affinities of human and rat NQO1 due to site‐specific phosphorylation

Abstract

HDXMS, hydrogen-deuterium exchange monitored by mass spectrometry; hNQO1, human NADP(H):quinone oxidoreductase 1; K d , dissociation constant; k prot , second-order rate constant for proteolysis; MD, molecular dynamics; NQO1 apo , ligand-free NQO1; NQO1 dic , FAD-dicoumarol-bound NQO1; NQO1 holo , FAD-bound NQO1; NTD, N-terminal domain; rNQO1, rat NADP(H):quinone oxidoreductase 1; TCS, thermolysin cleavage site; ΔΔG, change in the free energy change between two states (e.g. mutant and WT protein).