Sayğı Tuba Kuşman, Pınar Evren Gazel, Taştekil Ilgaz, Sarıca Pemra Özbek, Topçu Gülaçtı, Türkoğlu Emir Alper
Medicinal and Aromatic Plants Program, Hamidiye Vocational School of Health Services, University of Health Sciences Turkey, 34668, İstanbul, Türkiye.
Faculty of Pharmacy, University of Health Sciences Turkey, 34668, İstanbul, Türkiye.
Chem Biodivers. 2025 Feb;22(2):e202401470. doi: 10.1002/cbdv.202401470. Epub 2024 Nov 9.
In this study, the binding behavior of β-sitosterol with lysozyme (L) was elucidated by surface plasmon resonance (SPR), computational molecular docking and molecular dynamics simulation studies. Chicken egg white lysozyme (CEWL) served as a model protein. Tri-N-acetylchitotriose (NAG) was used in the redocking experiments to generate precise binding location of the protein. β-sitosterol displayed a slightly better binding energy (-6.68±0.04 kcal/mol) compared to NAG. Further molecular dynamics simulations and MMPBSA analysis revealed that residues Glu35, Gln57-Asn59, Trp62, Ile98, Ala107 and Trp108 contribute to the binding energy. Then, 2.5 mg/mL CEWL, 1X PBS buffer (pH 7.4) as running and coupling buffers, 30 μL/min as flow rate were applied for SPR analysis. Serial β-sitosterol injections (20-150 μM) were performed through SPR sensor surface. According to SPR binding study, K value for β-sitosterol-CEWL binding interaction was calculated as 71.34±9.79 μM. The results could provide essential knowledge for nutrition, pharmaceutical science, and oral biology.
在本研究中，通过表面等离子体共振（SPR）、计算分子对接和分子动力学模拟研究阐明了β-谷甾醇与溶菌酶（L）的结合行为。鸡卵清溶菌酶（CEWL）作为模型蛋白。在重新对接实验中使用三-N-乙酰壳三糖（NAG）来确定蛋白质的精确结合位置。与NAG相比，β-谷甾醇显示出稍好的结合能（-6.68±0.04 kcal/mol）。进一步的分子动力学模拟和MMPBSA分析表明，Glu35、Gln57-Asn59、Trp62、Ile98、Ala107和Trp108残基对结合能有贡献。然后，将2.5 mg/mL CEWL、1X PBS缓冲液（pH 7.4）用作运行和偶联缓冲液，流速为30 μL/min进行SPR分析。通过SPR传感器表面进行系列β-谷甾醇注射（20-150 μM）。根据SPR结合研究，计算出β-谷甾醇-CEWL结合相互作用的K值为71.34±9.79 μM。这些结果可为营养、制药科学和口腔生物学提供重要知识。
