Department of Physical Chemistry, University of Granada, Av. Fuentenueva S/N, 18071 Granada, Spain.
Int J Biol Macromol. 2018 Oct 15;118(Pt B):1912-1923. doi: 10.1016/j.ijbiomac.2018.07.051. Epub 2018 Jul 18.
Once whole-genome sequencing has reached the clinical practice, a main challenge ahead is the high-throughput and accurate prediction of the pathogenicity of genetic variants. However, current prediction tools do not consider explicitly a well-known property of disease-causing mutations: their ability to affect multiple functional sites distant in the protein structure. Here we carried out an extensive biophysical characterization of fourteen mutant variants at two cancer-associated sites of the enzyme NQO1, a paradigm of multi-functional protein. We showed that the magnitude of destabilizing effects, their molecular origins (structural vs. dynamic) and their efficient propagation through the protein structure gradually led to functional perturbations at different sites. Modulation of these structural perturbations also led to switches between molecular phenotypes. Our work supports that experimental and computational perturbation analyses would improve our understanding of the molecular basis of many loss-of-function genetic diseases as well as our ability to accurately predict the pathogenicity of genetic variants in a high-throughput fashion.
一旦全基因组测序进入临床实践，摆在面前的一个主要挑战是高通量、准确地预测遗传变异的致病性。然而，目前的预测工具并没有明确考虑到致病突变的一个众所周知的特性：它们能够影响蛋白质结构中远距离的多个功能位点。在这里，我们对两种癌症相关酶 NQO1 的两个位点的 14 个突变变体进行了广泛的生物物理特性分析，该酶是多功能蛋白的典范。我们表明，失稳效应的幅度、其分子起源（结构与动态）及其在蛋白质结构中的有效传播，逐渐导致不同位点的功能紊乱。对这些结构扰动的调节也导致了分子表型之间的转换。我们的工作支持这样的观点，即实验和计算扰动分析将有助于我们更好地理解许多功能丧失性遗传疾病的分子基础，以及我们以高通量方式准确预测遗传变异致病性的能力。
