The catalytic cycle of the antioxidant and cancer-associated human NQO1 enzyme: Hydride transfer, conformational dynamics and functional cooperativity

Anoz-Carbonell, E. (Universidad de Zaragoza) ; Timson, D.J. ; Pey, A.L. ; Medina, M. (Universidad de Zaragoza)

Resumen: Human NQO1 [NAD(H):quinone oxidoreductase 1] is a multi-functional and stress-inducible dimeric protein involved in the antioxidant defense, the activation of cancer prodrugs and the stabilization of oncosuppressors. Despite its roles in human diseases, such as cancer and neurological disorders, a detailed characterization of its enzymatic cycle is still lacking. In this work, we provide a comprehensive analysis of the NQO1 catalytic cycle using rapid mixing techniques, including multiwavelength and spectral deconvolution studies, kinetic modeling and temperature-dependent kinetic isotope effects (KIEs). Our results systematically support the existence of two pathways for hydride transfer throughout the NQO1 catalytic cycle, likely reflecting that the two active sites in the dimer catalyze two-electron reduction with different rates, consistent with the cooperative binding of inhibitors such as dicoumarol. This negative cooperativity in NQO1 redox activity represents a sort of half-of-sites activity. Analysis of KIEs and their temperature dependence also show significantly different contributions from quantum tunneling, structural dynamics and reorganizations to catalysis at the two active sites. Our work will improve our understanding of the effects of cancer-associated single amino acid variants and post-translational modifications in this protein of high relevance in cancer progression and treatment.

Idioma: Inglés

DOI: 10.3390/antiox9090772

Año: 2020

Publicado en: Antioxidants 9, 9 (2020), 772 [1-22]
ISSN: 2076-3921

Factor impacto JCR: 6.312 (2020)

Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 60 / 297 = 0.202 (2020) - Q1 - T1
Categ. JCR: FOOD SCIENCE & TECHNOLOGY rank: 11 / 144 = 0.076 (2020) - Q1 - T1
Categ. JCR: CHEMISTRY, MEDICINAL rank: 6 / 63 = 0.095 (2020) - Q1 - T1

Factor impacto SCIMAGO: 1.066 - Biochemistry (Q2) - Cell Biology (Q2) - Physiology (Q2) - Molecular Biology (Q2) - Clinical Biochemistry (Q2)

Financiación: info:eu-repo/grantAgreement/ES/DGA-FEDER/E35-20R
Financiación: info:eu-repo/grantAgreement/ES/MCIU-ERDF/RTI2018-096246-B-I00
Financiación: info:eu-repo/grantAgreement/ES/MICINN-AEI/PID2019-103901GB-I00

Tipo y forma: Article (Published version)

Área (Departamento): Área Bioquímica y Biolog.Mole. (Dpto. Bioq.Biolog.Mol. Celular)