@misc{10481/79025, year = {2022}, month = {11}, url = {https://hdl.handle.net/10481/79025}, abstract = {Recent advances in DNA sequencing technologies are revealing a large individual variability of the human genome. Our capacity to establish genotype-phenotype correlations in such large-scale is, however, limited. This task is particularly challenging due to the multifunctional nature of many proteins. Here we describe an extensive analysis of the stability and function of naturally-occurring variants (found in the COSMIC and gnomAD databases) of the cancer-associated human NAD(P)H:quinone oxidoreductase 1 (NQO1). First, we performed in silico saturation mutagenesis studies (>5,000 substitutions) aimed to identify regions in NQO1 important for stability and function. We then experimentally characterized twenty-two naturally-occurring variants in terms of protein levels during bacterial expression, solubility, thermal stability, and coenzyme binding. These studies showed a good overall correlation between experimental analysis and computational predictions; also the magnitude of the effects of the substitutions are similarly distributed in variants from the COSMIC and gnomAD databases. Outliers in these experimental-computational genotype-phenotype correlations remain, and we discuss these on the grounds and limitations of our approaches. Our work represents a further step to characterize the mutational landscape of NQO1 in the human genome and may help to improve high-throughput in silico tools for genotype-phenotype correlations in this multifunctional protein associated with disease.}, organization = {ERDF/Spanish Ministry of Science, Innovation and Universities-State Research Agency}, organization = {Junta de Andalucia RTI 2018-096246-B-I00}, organization = {ERDF/Counseling of Economic transformation, Industry, Knowledge and Universities P18-RT-2413}, organization = {Comunidad Valenciana B-BIO-84-UGR20}, organization = {Novo Nordisk Foundation}, organization = {Novocure Limited CIAICO/2021/135 NNF18OC0033950}, publisher = {Frontiers}, keywords = {Protein function}, keywords = {Protein stability}, keywords = {Genotype-phenotype correlations}, keywords = {Computational prediction}, keywords = {Sequence conservation}, title = {Effect of naturally-occurring mutations on the stability and function of cancer-associated NQO1: Comparison of experiments and computation}, doi = {10.3389/fmolb.2022.1063620}, author = {Pacheco García, Juan Luis and Tienne Matos, Kelly and Pey Rodríguez, Ángel Luis}, }