Effect of naturally-occurring mutations on the stability and function of cancer-associated NQO1: Comparison of experiments and computation
Research output: Contribution to journal › Journal article › Research › peer-review
Documents
- Fulltext
Final published version, 3.82 MB, PDF document
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.
Original language | English |
---|---|
Article number | 1063620 |
Journal | Frontiers in Molecular Biosciences |
Volume | 9 |
Number of pages | 17 |
ISSN | 2296-889X |
DOIs | |
Publication status | Published - 2022 |
- protein function, protein stability, genotype-phenotype correlations, computational prediction, sequence conservation, MICE LEADS, BINDING, OXIDOREDUCTASE-1, SUSCEPTIBILITY, RECOGNITION, DEGRADATION, DISRUPTION, VARIANTS, NAD(P)H, GENE
Research areas
ID: 329877403