Journal
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE
Volume 1842, Issue 11, Pages 2163-2173Publisher
ELSEVIER
DOI: 10.1016/j.bbadis.2014.08.011
Keywords
NAD(P)H quinone oxidoreductase 1; Cancer-associated polymorphism; NQO1*2; NQO1*3; Kinetic instability
Funding
- MINECO [CSD2009-00088, BIO2012-34937, RYC2009-04147]
- Junta de Andalucia [CTS-11-07187]
- FEDER Funds
- Department of Employment and Learning, Northern Ireland (DELNI)
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NAD(P) H quinone oxidoreductase 1 is involved in antioxidant defence and protection from cancer, stabilizing the apoptosis regulator p53 towards degradation. Here, we studied the enzymological, biochemical and biophysical properties of two cancer-associated variants (p.R139W and p.P1875). Both variants (especially p.1875) have lower thermal stability and greater susceptibility to proteolysis compared to the wild-type. p.P1875 also has reduced activity due to a lower binding affinity for the FAD cofactor as assessed by activity measurements and direct titrations. Native gel electrophoresis and dynamic light scattering also suggest that p.P1875 has a higher tendency to populate unfolded states under native conditions. Detailed thermal stability studies showed that all variants irreversibly denature causing dimer dissociation, while addition of FAD restores the stability of the polymorphic forms to wild-type levels. The kinetic destabilization induced by polymorphisms as well as the kinetic protection exerted by FAD was confirmed by measuring denaturation kinetics at temperatures close to physiological. Our data suggest that the main molecular mechanisms associated with these cancer-related variants are their low binding affinity for FAD and/or kinetic instability. Thus, pharmacological chaperones may be useful in the treatment of patients bearing these polymorphisms. (C) 2014 Elsevier B.V. All rights reserved.
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