Conformational dynamics is key to understanding loss-of-function of NQO1 cancer-associated polymorphisms and its correction by pharmacological ligands
0301 basic medicine
Dicumarol
Binding Sites
Protein Conformation
500
C700 Molecular Biology
Crystallography, X-Ray
HCT116 Cells
Polymorphism, Single Nucleotide
Article
3. Good health
03 medical and health sciences
Neoplasms
Biophysics and Biochemistry
Enzyme Stability
Flavin-Adenine Dinucleotide
NAD(P)H Dehydrogenase (Quinone)
Humans
Caco-2 Cells
Protein Multimerization
HeLa Cells
Protein Binding
DOI:
10.1038/srep20331
Publication Date:
2016-02-03T10:43:53Z
AUTHORS (8)
ABSTRACT
AbstractProtein 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.
SUPPLEMENTAL MATERIAL
Coming soon ....
REFERENCES (48)
CITATIONS (36)
EXTERNAL LINKS
PlumX Metrics
RECOMMENDATIONS
FAIR ASSESSMENT
Coming soon ....
JUPYTER LAB
Coming soon ....