Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations

Pacheco-Garcia, Juan Luis ; Loginov, Dmitry S. ; Anoz-Carbonell, Ernesto ; Vankova, Pavla ; Palomino-Morales, Rogelio ; Salido, Eduardo ; Man, Petr ; Medina, Milagros (Universidad de Zaragoza) ; Naganathan, Athi N. ; Pey, Angel L.

Resumen: Allosterism is a common phenomenon in protein biochemistry that allows rapid regulation of protein stability; dynamics and function. However, the mechanisms by which allosterism occurs (by mutations or post-translational modifications (PTMs)) may be complex, particularly due to long-range propagation of the perturbation across protein structures. In this work, we have investigated allosteric communication in the multifunctional, cancer-related and antioxidant protein NQO1 by mutating several fully buried leucine residues (L7, L10 and L30) to smaller residues (V, A and G) at sites in the N-terminal domain. In almost all cases, mutated residues were not close to the FAD or the active site. Mutations L\u2192G strongly compromised conformational stability and solubility, and L30A and L30V also notably decreased solubility. The mutation L10A, closer to the FAD binding site, severely decreased FAD binding affinity (\u224820 fold vs. WT) through long-range and context-dependent effects. Using a combination of experimental and computational analyses, we show that most of the effects are found in the apo state of the protein, in contrast to other common polymorphisms and PTMs previously characterized in NQO1. The integrated study presented here is a first step towards a detailed structural-functional mapping of the mutational landscape of NQO1, a multifunctional and redox signaling protein of high biomedical relevance.

Idioma: Inglés

DOI: 10.3390/antiox11061110

Año: 2022

Publicado en: Antioxidants 11, 6 (2022), 1110
ISSN: 2076-3921

Factor impacto JCR: 7.0 (2022)

Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 46 / 285 = 0.161 (2022) - Q1 - T1
Categ. JCR: FOOD SCIENCE & TECHNOLOGY rank: 13 / 142 = 0.092 (2022) - Q1 - T1
Categ. JCR: CHEMISTRY, MEDICINAL rank: 6 / 60 = 0.1 (2022) - Q1 - T1

Factor impacto CITESCORE: 8.8 - Biochemistry, Genetics and Molecular Biology (Q1)

Factor impacto SCIMAGO: 1.084 - Biochemistry (Q1) - Clinical Biochemistry (Q1) - Food Science (Q1) - Physiology (Q1) - Molecular Biology (Q2) - Cell Biology (Q2)

Financiación: info:eu-repo/grantAgreement/ES/DGA-FEDER/E35-20R
Financiación: info:eu-repo/grantAgreement/ES/ERDF-B-BIO-84-UGR20
Financiación: info:eu-repo/grantAgreement/EC/H2020/82383/EPIC-XS/EU
Financiación: info:eu-repo/grantAgreement/ES/JA-P18-RT-2413
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)