Department of Biology, Faculty of Science, Shahrekord University, Shahrekord, P. O. Box 115, Iran; Central Laboratory, Shahrekord University, Shahrekord, Iran.
Department of Biology, Faculty of Science, Shahrekord University, Shahrekord, P. O. Box 115, Iran; Central Laboratory, Shahrekord University, Shahrekord, Iran.
Int J Biol Macromol. 2023 Dec 31;253(Pt 1):126605. doi: 10.1016/j.ijbiomac.2023.126605. Epub 2023 Sep 3.
As an inevitable parameter in the description of enzyme properties, the investigation of enzyme-ligand interactions has attracted a lot of attention. Alpha-Chymotrypsin (α-Chy) is essential for protein digestion and plays an important role in human health. Naringenin (NAG) as a potent antioxidant has recently been applied in the pharmaceutical industry. Using multispectral methods and computational simulation techniques, the binding strength of NAG to α-Chy was investigated in this research. UV-vis and fluorescence quenching data showed significant spectral changes upon binding of NAG to α-Chy. As demonstrated by fluorescence techniques, NAG could employ a static quenching process to decrease the intrinsic fluorescence of α-Chy. Both circular dichroism (CD) and FTIR spectroscopic analyses revealed that binding of NAG to α-Chy caused more flexible conformation. The slight increases in RMSD (0.06 nm) were observed for the NAG-(α-Chy) compound was supported by the results of thermal stability data. Docking computation confirmed that hydrogen and Van der Waals interactions are the important forces, which is in exact agreement with thermodynamics studies. Kinetic analysis of the enzyme showed an increase in activity, which was consistent, with the MD simulation results. The findings from the in-silico studies were in complete agreement with the experimental results.
作为酶性质描述中不可避免的参数，对酶-配体相互作用的研究引起了广泛关注。糜蛋白酶（α-Chy）是蛋白质消化所必需的，在人类健康中起着重要作用。柚皮素（NAG）作为一种有效的抗氧化剂，最近已在制药行业中得到应用。本研究采用多光谱方法和计算模拟技术，研究了 NAG 与 α-Chy 的结合强度。紫外-可见和荧光猝灭数据表明，NAG 与 α-Chy 结合后会发生显著的光谱变化。荧光技术表明，NAG 可以通过静态猝灭过程降低α-Chy 的本征荧光。圆二色性（CD）和傅里叶变换红外（FTIR）光谱分析表明，NAG 与 α-Chy 的结合导致其构象更加灵活。由热稳定性数据可知，NAG-（α-Chy）化合物的 RMSD（0.06nm）略有增加。对接计算结果证实，氢键和范德华相互作用是重要的作用力，这与热力学研究结果完全一致。对酶的动力学分析表明活性增加，这与 MD 模拟结果一致。计算机模拟研究的结果与实验结果完全一致。
