Structural basis of the pleiotropic and specific phenotypic consequences of missense mutations in the multifunctional NAD(P)H:quinone oxidoreductase 1 and their pharmacological rescue

• Published in: Redox biology Vol. 46; p. 102112
• Main Authors: Pacheco-Garcia, Juan Luis, Anoz-Carbonell, Ernesto, Vankova, Pavla, Kannan, Adithi, Palomino-Morales, Rogelio, Mesa-Torres, Noel, Salido, Eduardo, Man, Petr, Medina, Milagros, Naganathan, Athi N., Pey, Angel L.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.10.2021; Elsevier
• Subjects: Biochemistry; Biochemistry, Molecular Biology; Biophysics; Disease-causing mutation; Flavin-Adenine Dinucleotide - metabolism; Flavoprotein; Humans; Life Sciences; Ligand binding; Multifunctional protein; Mutation, Missense; NAD; NAD(P)H Dehydrogenase (Quinone) - genetics; NAD(P)H Dehydrogenase (Quinone) - metabolism; NQO1; Post-translational modification; Protein Binding; Quinones; Research Paper; Structural Biology; Post-translational modification; Multifunctional protein; Ligand binding; NQO1; Disease-causing mutation; Flavoprotein; antioxidant enzyme; Flavoprotein Multifunctional protein Ligand binding Disease-causing mutation Post-translational modification NQO1; enzyme kinetic analysis; hydride transfer; conformational dynamics; quantum tunneling; cancer; functional cooperativity; oxidoreductase; antioxidant response; kinetic isotope effects
• ISSN: 2213-2317; 2213-2317
• DOI: 10.1016/j.redox.2021.102112

The multifunctional nature of human flavoproteins is critically linked to their ability to populate multiple conformational states. Ligand binding, post-translational modifications and disease-associated mutations can reshape this functional landscape, although the structure-function relationships of these effects are not well understood. Herein, we characterized the structural and functional consequences of two mutations (the cancer-associated P187S and the phosphomimetic S82D) on different ligation states which are relevant to flavin binding, intracellular stability and catalysis of the disease-associated NQO1 flavoprotein. We found that these mutations affected the stability locally and their effects propagated differently through the protein structure depending both on the nature of the mutation and the ligand bound, showing directional preference from the mutated site and leading to specific phenotypic manifestations in different functional traits (FAD binding, catalysis and inhibition, intracellular stability and pharmacological response to ligands). Our study thus supports that pleitropic effects of disease-causing mutations and phosphorylation events on human flavoproteins may be caused by long-range structural propagation of stability effects to different functional sites that depend on the ligation-state and site-specific perturbations. Our approach can be of general application to investigate these pleiotropic effects at the flavoproteome scale in the absence of high-resolution structural models. [Display omitted] •Flavoprotein function is affected by ligands, mutations and posttranslational modifications.•The structural and energetic basis of these effects are not well understood.•We carried out extensive structure-function studies on human NQO1 WT and two mutants.•Mutations cause long-range effects on different functional sites depending on the bound ligand.•Phenotypic consequences are rationalized by these extensive structure-function studies.