Departamento de Química Física, Facultad de Ciencias, Universidad de Granada, Spain.
Institute of Microbiology of the Czech Academy of Sciences, BioCeV, Praha, Czech Republic.
FEBS Lett. 2022 Jan;596(1):29-41. doi: 10.1002/1873-3468.14238. Epub 2021 Dec 9.
The phosphomimetic mutation S82D in the cancer-associated, FAD-dependent human NADP(H):quinone oxidoreductase 1 (hNQO1) causes a decrease in flavin-adenine dinucleotide-binding affinity and intracellular stability. We test in this work whether the evolutionarily recent neutral mutation R80H in the vicinity of S82 may alter the strong functional effects of S82 phosphorylation through electrostatic interactions. We show using biophysical and bioinformatic analyses that the reverse mutation H80R prevents the effects of S82D phosphorylation on hNQO1 by modulating the local stability. Consistently, in rat NQO1 (rNQO1) which contains R80, the effects of phosphorylation were milder, resembling the behaviour found in hNQO1 when this residue was humanized in rNQO1 (by the R80H mutation). Thus, apparently neutral and evolutionarily divergent mutations may determine the functional response of mammalian orthologues towards phosphorylation.
癌症相关的 FAD 依赖性人 NADP(H)：醌氧化还原酶 1 (hNQO1) 中的磷酸模拟突变 S82D 导致黄素腺嘌呤二核苷酸结合亲和力和细胞内稳定性降低。在这项工作中，我们测试了附近的进化上新的中性突变 R80H 是否可以通过静电相互作用改变 S82 磷酸化的强功能效应。我们通过生物物理和生物信息学分析表明，反向突变 H80R 通过调节局部稳定性来阻止 S82D 磷酸化对 hNQO1 的影响。一致地，在含有 R80 的大鼠 NQO1（rNQO1）中，磷酸化的影响较轻，类似于在 rNQO1 中该残基被人源化（通过 R80H 突变）时发现的 hNQO1 的行为。因此，显然中性和进化上不同的突变可能决定了哺乳动物同源物对磷酸化的功能反应。
