Marycleopha Manka, Johnson Jennifer, Singh Abhishek, Kumar Satish
Laboratory of Forensic Biology and Biotechnology, School of Forensic Science, National Forensic Science University, Gandhinagar, Gujarat, 382007, India.
National Forensic Sciences University, Goa Campus, Ponda, Goa, 403401, India.
Forensic Toxicol. 2024 Dec 21. doi: 10.1007/s11419-024-00709-w.
This study examines the interaction between benzoylmesaconine (BMA) and hen egg white lysozyme (HEWL) under various physiological conditions, aiming to determine how BMA affects the HEWL's structure and function.
Several analytical techniques were used, including tryptophan assay, light scattering, thioflavin T (ThT)-binding assay, dynamic light scattering, 8-anilino-1-naphthalenesulfonic acid (ANS)-binding assay, circular dichroism (CD) spectroscopy, enzyme activity assay, and molecular docking.
The tryptophan assay displayed a concentration-dependent decrease in tryptophan fluorescence, showing an interaction between BMA and HEWL. Light scattering and ThT-binding assays confirmed increased protein aggregation and amyloid fibril formation, while the ANS-binding assay demonstrated altered exposed hydrophobic regions, implying structural changes. CD spectroscopy showed a reduction in α-helix content, indicating conformational alterations, and enzyme activity assays showed a loss of lytic function due to structural distortion. Finally, molecular docking identified significant bonds and hydrophobic interactions between BMA and HEWL residues.
BMA binding induces structural changes in proteins, forming small oligomers and amyloid fibrils that decrease HEWL enzymatic activity and disrupt functional integrity.
本研究考察了在不同生理条件下苯甲酰美沙可宁（BMA）与鸡蛋清溶菌酶（HEWL）之间的相互作用，旨在确定BMA如何影响HEWL的结构和功能。
使用了多种分析技术，包括色氨酸测定、光散射、硫黄素T（ThT）结合测定、动态光散射、8-苯胺基-1-萘磺酸（ANS）结合测定、圆二色（CD）光谱、酶活性测定和分子对接。
色氨酸测定显示色氨酸荧光呈浓度依赖性降低，表明BMA与HEWL之间存在相互作用。光散射和ThT结合测定证实蛋白质聚集增加和淀粉样纤维形成，而ANS结合测定表明暴露的疏水区域发生改变，这意味着结构发生了变化。CD光谱显示α-螺旋含量降低，表明构象改变，酶活性测定显示由于结构扭曲导致溶菌功能丧失。最后，分子对接确定了BMA与HEWL残基之间的重要键和疏水相互作用。
BMA结合诱导蛋白质结构变化，形成小寡聚体和淀粉样纤维，降低HEWL酶活性并破坏功能完整性。
