Department of Integrative Biology, School of Bio Sciences and Technology, VIT, Vellore, Tamil Nadu, India.
Faculty of Computing and Information Technology, King Abdulaziz University, Rabigh, Saudi Arabia.
Adv Protein Chem Struct Biol. 2019;114:341-407. doi: 10.1016/bs.apcsb.2018.10.009. Epub 2018 Dec 18.
Fabry's disease (FD) is the second most commonly occurring lysosomal storage disorders (LSDs). The mutations in α-galactosidase A (GLA) protein were widely found to be causative for the Fabry's disease. These mutations result in alternate splicing methods that affect the stability and function of the protein. The mutations near the active site of the protein results in protein misfolding. In this study, we have retrieved the missense mutation data from the three public databases (NCBI, UniProt, and HGMD). We used multiple in silico tools to predict the pathogenicity and stability of these mutations. Mutations in the active sites (D92Y, C142Y, D170V, and D266N) of the protein were screened for the phenotyping analysis using SNPeffect 4.0. Mutant D92Y was predicted to increase the amyloid propensity as well as severely reduce the protein stability and the remaining mutations showed no significant results by SNPeffect 4.0. Protein dynamics simulations (PDS) were performed to understand the behavior of the proteins due to the mutations. The simulation results showed that the D92Y mutant was more severe (higher deviation, loss of intramolecular hydrogen bonds, and lower compactness) than the other protein mutants (C142Y, D170V, and D266N). Further, the action of pharmacological chaperone 1-deoxygalactonojirimycin (DGJ) over the severe mutation was studied using the molecular docking analysis. Chaperone DGJ, an iminosugar plays a convincing role in repairing the misfolded protein and helps the protein to achieve its normal function. From the molecular docking analysis, we observed that both the native protein and protein with D92Y mutation followed similar interaction patterns. Further, the docked complexes (native-DGJ and mutant-DGJ) were subjected to PDS analysis. From the simulation analysis, we observed that DGJ had shown the better effect on the protein with the D92Y mutation. This elucidates that DGJ can still be used as a promising chaperone to treat the FD caused by mutations of GLA protein.
法布里病（FD）是第二常见的溶酶体贮积症（LSDs）。α-半乳糖苷酶 A（GLA）蛋白的突变被广泛认为是法布里病的病因。这些突变导致替代剪接方式，影响蛋白质的稳定性和功能。蛋白质活性部位附近的突变导致蛋白质错误折叠。在这项研究中，我们从三个公共数据库（NCBI、UniProt 和 HGMD）中检索了错义突变数据。我们使用多种计算工具来预测这些突变的致病性和稳定性。使用 SNPeffect 4.0 对蛋白质活性部位（D92Y、C142Y、D170V 和 D266N）的突变进行表型分析筛选。突变 D92Y 被预测为增加淀粉样倾向，并严重降低蛋白质稳定性，而其余突变则未显示出 SNPeffect 4.0 的显著结果。进行蛋白质动力学模拟（PDS）以了解由于突变导致的蛋白质行为。模拟结果表明，突变 D92Y 比其他蛋白质突变体（C142Y、D170V 和 D266N）更严重（更大的偏差、丧失分子内氢键和更低的紧凑性）。此外，使用分子对接分析研究了药理伴侣 1-脱氧半乳糖基氮己吡喃糖苷（DGJ）对严重突变的作用。伴侣 DGJ，一种亚氨基糖，在修复错误折叠的蛋白质方面发挥了令人信服的作用，并帮助蛋白质实现其正常功能。从分子对接分析中，我们观察到天然蛋白和 D92Y 突变蛋白都遵循类似的相互作用模式。此外，对接复合物（天然-DGJ 和突变-DGJ）进行了 PDS 分析。从模拟分析中，我们观察到 DGJ 对 D92Y 突变蛋白的效果更好。这表明 DGJ 仍然可以用作有前途的伴侣来治疗由 GLA 蛋白突变引起的 FD。
