title: Conformational dynamics is key to understanding loss-of-function of NQO1 cancer-associated polymorphisms and its correction by pharmacological ligands.
abstract: Protein dynamics is essential to understand protein function and stability, even though is rarely investigated as the origin of loss-of-function due to genetic variations. Here, we use biochemical, biophysical, cell and computational biology tools to study two loss-of-function and cancer-associated polymorphisms (p.R139W and p.P187S) in human NAD(P)H quinone oxidoreductase 1 (NQO1), a FAD-dependent enzyme which activates cancer pro-drugs and stabilizes several oncosuppressors. We show that p.P187S strongly destabilizes the NQO1 dimer in vitro and increases the flexibility of the C-terminal domain, while a combination of FAD and the inhibitor dicoumarol overcome these alterations. Additionally, changes in global stability due to polymorphisms and ligand binding are linked to the dynamics of the dimer interface, whereas the low activity and affinity for FAD in p.P187S is caused by increased fluctuations at the FAD binding site. Importantly, NQO1 steady-state protein levels in cell cultures correlate primarily with the dynamics of the C-terminal domain, supporting a directional preference in NQO1 proteasomal degradation and the use of ligands binding to this domain to stabilize p.P187S in vivo. In conclusion, protein dynamics are fundamental to understanding loss-of-function in p.P187S, and to develop new pharmacological therapies to rescue this function.
authors.affiliation: Department of Physical Chemistry, Faculty of Sciences, University of Granada, Granada, Spain.
authors.forename: Encarnación
authors.initials: E
authors.lastname: Medina-Carmona
authors.affiliation: Department of Biochemistry and Molecular Biology I, Faculty of Sciences, University of Granada, Granada, Spain.
authors.forename: Rogelio J
authors.initials: RJ
authors.lastname: Palomino-Morales
authors.affiliation: Centre for Molecular Informatics, Department of Chemistry, University of Cambridge, Cambridge, UK.
authors.forename: Julian E
authors.initials: JE
authors.lastname: Fuchs
authors.affiliation: Department of Physical Chemistry, Faculty of Sciences, University of Granada, Granada, Spain.
authors.forename: Esperanza
authors.initials: E
authors.lastname: Padín-Gonzalez
authors.affiliation: Department of Physical Chemistry, Faculty of Sciences, University of Granada, Granada, Spain.
authors.forename: Noel
authors.initials: N
authors.lastname: Mesa-Torres
authors.affiliation: Hospital Universitario de Canarias, Tenerife, Spain.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Salido
authors.affiliation: School of Biological Sciences, Queen´s University Belfast, Belfast, UK.|School of Pharmacy and Biomolecular Sciences, The University of Brighton, Brighton, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Timson
authors.affiliation: Department of Physical Chemistry, Faculty of Sciences, University of Granada, Granada, Spain.
authors.forename: Angel L
authors.initials: AL
authors.lastname: Pey
mesh_terms: D001665:Binding Sites / Q000187:drug effects; D018938:Caco-2 Cells; D018360:Crystallography, X-Ray; D001728:Dicumarol / Q000494:pharmacology*; D004795:Enzyme Stability / Q000187:drug effects; D005182:Flavin-Adenine Dinucleotide / Q000494:pharmacology*; D045325:HCT116 Cells; D006367:HeLa Cells; D006801:Humans; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D009369:Neoplasms / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide; D011485:Protein Binding / Q000187:drug effects; D011487:Protein Conformation / Q000187:drug effects; D055503:Protein Multimerization
chemical_list: D005182:Flavin-Adenine Dinucleotide; D001728:Dicumarol; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human
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