Zebrafish Genetics Laboratory, Department of Genetic Engineering, School of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Chengalpattu, Tamil Nadu, India.
Department of Biotechnology, Marudhar Kesari Jain College for Women, Vaniyambadi, Vellore, Tamil Nadu, India.
J Biomol Struct Dyn. 2022 Feb;40(3):1260-1272. doi: 10.1080/07391102.2020.1824812. Epub 2020 Sep 24.
Fumarate hydratase (FH), one of the members of TCA cycle, acts as a catalyte for the synthesis of malate from fumarate. FH has been proposed to play as a tumour suppressor leading to the pathogenicity of leiomyomas, renal cell carcinoma and paraganglioma. Mutations in the active site of FH lead to alteration in the protein structure. Similarly, binding of several chemical inhibitors to the active site also leads to the disruption of protein structural integrity thereby leading to protein dysfunction. Therefore, in order to address this mechanism leading to cancer, the binding efficiency of potential human FH inhibitor citrate to zebrafish fh has been extensively analysed in this study by molecular docking and simulation experiments followed by quantification of fumarate hydratase enzyme activity to validate and confirm the findings. Molecular docking revealed stronger interaction of zebrafish fh protein with inhibitor citrate when compared to natural substrate fumarate. Study on the dynamics of docked structures further confirmed that citrate was found to possess more binding affinity than fumarate. biochemical analysis also revealed concentration dependent potential inhibitory effect of citrate on zebrafish fh, thus confirming the findings of the experiments.Communicated by Ramaswamy H. Sarma.
延胡索酸水合酶（FH）是三羧酸循环的成员之一，作为催化延胡索酸转化为苹果酸的酶。FH 被认为是一种肿瘤抑制因子，导致平滑肌瘤、肾细胞癌和副神经节瘤的发病。FH 活性部位的突变导致蛋白结构的改变。同样，几种化学抑制剂结合到活性部位也会导致蛋白结构完整性的破坏，从而导致蛋白功能障碍。因此，为了解决导致癌症的这种机制，本研究通过分子对接和模拟实验，广泛分析了潜在的人 FH 抑制剂柠檬酸盐与斑马鱼 fh 的结合效率，并通过测定延胡索酸水合酶活性对结果进行验证和确认。分子对接显示，与天然底物延胡索酸相比，斑马鱼 fh 蛋白与抑制剂柠檬酸盐的相互作用更强。对接结构动力学的研究进一步证实，柠檬酸盐的结合亲和力大于延胡索酸。生化分析还揭示了柠檬酸盐对斑马鱼 fh 的浓度依赖性潜在抑制作用，从而证实了实验结果。由 Ramaswamy H. Sarma 传达。
