Citral Inhibition of Human Salivary Aldehyde Dehydrogenase

Human salivary aldehyde dehydrogenase (hsALDH) protects us from the toxic effect of aldehydes. It has both diagnostic and therapeutic importance. Citral possesses many biological and pharmacological properties. The aim of this work was to investigate the inhibitory effect and the mechanism of inhibition of citral on hsALDH. Citral inhibits the dehydrogenase activity of hsALDH. It decreased the substrate affinity and to a lesser extent, the catalytic efficiency of hsALDH. Citral showed linear mixed-type inhibition with a higher tendency of competitive behavior with little, but significant, non-competitive inhibition. The nucleophilicity of active site Cys residue is not a significant contributing factor in the inhibition process. Citral shows uncompetitive inhibition towards the co-enzyme (NAD⁺). α-helix and β-sheet content of the enzyme were changed in presence of citral. Biophysical studies showed that citral quenches the intrinsic fluorescence of hsALDH in a static manner by forming complex with the enzyme. Molecular docking study showed that both the isomers of citral bind to the catalytic site of hsALDH interacting with few evolutionary preserved amino acid residues through multiple non-covalent interactions. Ligand efficiency metrics values indicate that citral is an efficient ligand for the enzyme in terms of its physicochemical and pharmacokinetic properties.