The induction of chirality in a ligand can be a powerful analytical tool for studying protein–ligand interactions. Here, we advanced by applying the technique to monitor the inversion of the induced circular dichroism (ICD) spectrum when ligands move between human and bovine serum albumin proteins (HSA and BSA). ICD experiments were performed using dimers of methyl vanillate (DVT) and vanillin (DVN). The sign and spectra shape were dependent on the albumin type. DVN presented a positive maximum in 312 nm when complexed with HSA and a negative one in BSA. It was possible to induce and follow the time-dependent displacement of the ligand from BSA (2.2 × 10⁶ M⁻¹) to HSA (6.6 × 10⁵ M⁻¹) via ICD inversion. The Molecular Mechanics Generalized Born Surface Area approach was used to calculate the binding free energy of the conformers, and a dissociation pathway for each system was proposed using Umbrella Sampling calculations. Four energy minima dihedral angle conformers were identified, and the corresponding CD spectra were calculated using the quantum chemistry approach. Then, weighted spectra for the conformationally accessible conformers were obtained based on each conformer's Boltzmann probability distribution. In conclusion, the methodology described in the manuscript might be helpful in monitoring the movement of ligands between proteins that they bind.

配体中手性的诱导可以成为研究蛋白质-配体相互作用的有力分析工具。在这里，我们通过应用该技术来监测配体在人和牛血清白蛋白（HSA 和 BSA）之间移动时的诱导圆二色性 (ICD) 光谱的反转。使用香草酸甲酯 (DVT) 和香草醛 (DVN) 的二聚体进行 ICD 实验。符号和光谱形状取决于白蛋白类型。当与 HSA 复合时，DVN 在 312 nm 处呈现正最大值，而在 BSA 中呈现负最大值。^{可以诱导和跟踪配体从 BSA (2.2 × 10 6}  M ^-1 ) 到 HSA (6.6 × 10 ⁵  M ^-1 )的时间依赖性置换) 通过 ICD 反转。分子力学广义玻恩表面积方法用于计算构象异构体的结合自由能，并使用伞形采样计算提出了每个系统的解离途径。确定了四个能量最小值二面角构象异构体，并使用量子化学方法计算了相应的 CD 光谱。然后，基于每个构象异构体的玻尔兹曼概率分布获得构象可接近构象异构体的加权光谱。总之，手稿中描述的方法可能有助于监测配体在它们结合的蛋白质之间的运动。