117263 20240319081016.0 doi 10.3390/antiox11061110 sideral 128988 ART-2022-128988 eng Pacheco-Garcia, Juan Luis Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations 2022 Access copy available to the general public Unrestricted 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. info:eu-repo/grantAgreement/ES/DGA-FEDER/E35-20R info:eu-repo/grantAgreement/ES/ERDF-B-BIO-84-UGR20 info:eu-repo/grantAgreement/EC/H2020/82383/EPIC-XS/EU This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 82383 EPIC-XS info:eu-repo/grantAgreement/ES/JA-P18-RT-2413 info:eu-repo/grantAgreement/ES/MCIU-ERDF/RTI2018-096246-B-I00 info:eu-repo/grantAgreement/ES/MICINN-AEI/PID2019-103901GB-I00 info:eu-repo/semantics/openAccess by http://creativecommons.org/licenses/by/3.0/es/ 7.0 2022 BIOCHEMISTRY & MOLECULAR BIOLOGY 46 / 285 = 0.161 2022 Q1 T1 FOOD SCIENCE & TECHNOLOGY 13 / 142 = 0.092 2022 Q1 T1 CHEMISTRY, MEDICINAL 6 / 60 = 0.1 2022 Q1 T1 1.084 2022 Biochemistry 2022 Q1 Clinical Biochemistry 2022 Q1 Food Science 2022 Q1 Physiology 2022 Q1 Molecular Biology 2022 Q2 Cell Biology 2022 Q2 8.8 2022 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Loginov, Dmitry S. Anoz-Carbonell, Ernesto (orcid)0000-0002-6649-9153 Vankova, Pavla Palomino-Morales, Rogelio Salido, Eduardo Man, Petr Medina, Milagros Universidad de Zaragoza (orcid)0000-0001-8743-0182 Naganathan, Athi N. Pey, Angel L. 1002 060 Universidad de Zaragoza Dpto. Bioq.Biolog.Mol. Celular Área Bioquímica y Biolog.Mole. 11, 6 (2022), 1110 Antioxidants Antioxidants 2076-3921 4370878 http://zaguan.unizar.es/record/117263/files/texto_completo.pdf Versión publicada 2740188 http://zaguan.unizar.es/record/117263/files/texto_completo.jpg?subformat=icon icon Versión publicada oai:zaguan.unizar.es:117263 articulos driver 2024-03-18-15:40:05 ARTICLE