Number of the records: 1
Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations
- 1.
SYSNO ASEP 0564248 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Allosteric Communication in the Multifunctional and Redox NQO1 Protein Studied by Cavity-Making Mutations Author(s) Pacheco-Garcia, J. L. (ES)
Loginov, Dmitry Sergej (MBU-M) RID
Anoz-Carbonell, E. (ES)
Vaňková, Pavla (MBU-M) ORCID
Palomino-Morales, R. (ES)
Salido, E. (ES)
Man, Petr (MBU-M) RID, ORCID
Medina, M. (ES)
Naganathan, A. N. (IN)
Pey, Angel L. (ES)Article number 1110 Source Title Antioxidants. - : MDPI
Roč. 11, č. 6 (2022)Number of pages 16 s. Language eng - English Country CH - Switzerland Keywords antioxidant defense ; flavoprotein ; FAD binding ; structural perturbation ; protein core ; allosterism ; cavity-making mutation Subject RIV CE - Biochemistry OECD category Biochemistry and molecular biology Subject RIV - cooperation Institute of Biotechnology R&D Projects ED1.1.00/02.0109 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Research Infrastructure CIISB II - 90127 - Masarykova univerzita Method of publishing Open access Institutional support MBU-M - RVO:61388971 ; BTO-N - RVO:86652036 UT WOS 000816602700001 EID SCOPUS 85131635705 DOI 10.3390/antiox11061110 Annotation 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> G 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 (approximate to 20 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. Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2023 Electronic address https://www.mdpi.com/2076-3921/11/6/1110
Number of the records: 1