000121158 001__ 121158
000121158 005__ 20241125101126.0
000121158 0247_ $$2doi$$a10.1111/febs.16677
000121158 0248_ $$2sideral$$a131629
000121158 037__ $$aART-2023-131629
000121158 041__ $$aeng
000121158 100__ $$aPacheco-García, Juan Luis
000121158 245__ $$aCounterintuitive structural and functional effects due to naturally occurring mutations targeting the active site of the disease-associated NQO1 enzyme*
000121158 260__ $$c2023
000121158 5060_ $$aAccess copy available to the general public$$fUnrestricted
000121158 5203_ $$aOur knowledge on the genetic diversity of the human genome is exponentially growing. However, our capacity to establish genotype–phenotype correlations on a large scale requires a combination of detailed experimental and computational work. This is a remarkable task in human proteins which are typically multifunctional and structurally complex. In addition, mutations often prevent the determination of mutant high-resolution structures by X-ray crystallography. We have characterized here the effects of five mutations in the active site of the disease-associated NQO1 protein, which are found either in cancer cell lines or in massive exome sequencing analysis in human population. Using a combination of H/D exchange, rapid-flow enzyme kinetics, binding energetics and conformational stability, we show that mutations in both sets may cause counterintuitive functional effects that are explained well by their effects on local stability regarding different functional features. Importantly, mutations predicted to be highly deleterious (even those affecting the same protein residue) may cause mild to catastrophic effects on protein function. These functional effects are not well explained by current predictive bioinformatic tools and evolutionary models that account for site conservation and physicochemical changes upon mutation. Our study also reinforces the notion that naturally occurring mutations not identified as disease-associated can be highly deleterious. Our approach, combining protein biophysics and structural biology tools, is readily accessible to broadly increase our understanding of genotype–phenotype correlations and to improve predictive computational tools aimed at distinguishing disease-prone against neutral missense variants in the human genome.
000121158 536__ $$9info:eu-repo/grantAgreement/ES/DGA-FEDER/E35-20R$$9info:eu-repo/grantAgreement/ES/ERDF-B-BIO-84-UGR20$$9info:eu-repo/grantAgreement/EC/H2020/731077/EU/European Network of Fourier-Transform Ion-Cyclotron-Resonance Mass Spectrometry Centers/EU_FT-ICR_MS$$9This project has received funding from the European Union’s Horizon 2020 research and innovation program under grant agreement No H2020 731077-EU_FT-ICR_MS$$9info:eu-repo/grantAgreement/ES/MCIU-ERDF/RTI2018-096246-B-I00$$9info:eu-repo/grantAgreement/ES/MICINN-AEI/PID2019-103901GB-I00
000121158 540__ $$9info:eu-repo/semantics/openAccess$$aby$$uhttp://creativecommons.org/licenses/by/3.0/es/
000121158 590__ $$a5.5$$b2023
000121158 592__ $$a2.003$$b2023
000121158 591__ $$aBIOCHEMISTRY & MOLECULAR BIOLOGY$$b58 / 313 = 0.185$$c2023$$dQ1$$eT1
000121158 593__ $$aBiochemistry$$c2023$$dQ1
000121158 593__ $$aMolecular Biology$$c2023$$dQ1
000121158 593__ $$aCell Biology$$c2023$$dQ1
000121158 594__ $$a11.7$$b2023
000121158 655_4 $$ainfo:eu-repo/semantics/article$$vinfo:eu-repo/semantics/publishedVersion
000121158 700__ $$aAnoz Carbonell, Ernesto
000121158 700__ $$aLoginov, Dmitry S.
000121158 700__ $$aKavan, Daniel
000121158 700__ $$aSalido, Eduardo
000121158 700__ $$aMan, Petr
000121158 700__ $$0(orcid)0000-0001-8743-0182$$aMedina, Milagros$$uUniversidad de Zaragoza
000121158 700__ $$aPey, Angel L.
000121158 7102_ $$11002$$2060$$aUniversidad de Zaragoza$$bDpto. Bioq.Biolog.Mol. Celular$$cÁrea Bioquímica y Biolog.Mole.
000121158 773__ $$g290, 7 (2023), 1855-1873$$pFEBS J.$$tFEBS Journal$$x1742-464X
000121158 8564_ $$s4431512$$uhttps://zaguan.unizar.es/record/121158/files/texto_completo.pdf$$yVersión publicada
000121158 8564_ $$s2578736$$uhttps://zaguan.unizar.es/record/121158/files/texto_completo.jpg?subformat=icon$$xicon$$yVersión publicada
000121158 909CO $$ooai:zaguan.unizar.es:121158$$particulos$$pdriver
000121158 951__ $$a2024-11-22-11:57:45
000121158 980__ $$aARTICLE