Quantitative analysis of molecular partition towards lipid membranes using surface plasmon resonance.
Quantitative analysis of molecular partition towards lipid membranes using surface plasmon resonance.
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DOI：
10.1038/srep45647
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发表时间：
2017-03-30
影响因子：
4.6
通讯作者：
Castanho MA
中科院分区：
文献类型：
--
作者：
Figueira TN;Freire JM;Cunha-Santos C;Heras M;Gonçalves J;Moscona A;Porotto M;Salomé Veiga A;Castanho MA
了解分子和脂质膜之间的相互作用是研究细胞和生物技术现象的基础。水介质和脂质膜之间的分配是许多生物分子和药物作用机制的关键。因此，通过适当和稳健的参数来量化膜分配是至关重要的。表面等离子体共振（SPR）是一种研究1：1化学计量相互作用的强大技术，但对脂膜分配数据的应用有限。我们已经开发并应用了一种新的数学模型SPR数据处理，使动力学和平衡分配常数的测定。该方法使用两个互补的拟合模型的关联和解离传感图数据。将抗体片段F63、HIV融合抑制剂恩夫韦肽和内源性药物kyotorphin对POPC膜获得的SPR分配数据与来自独立技术的数据进行比较。全面的动力学和分配模型被施加到膜相互作用的数据HRC 4，麻疹病毒进入抑制肽，揭示其增加的亲和力，并保留在富含胆固醇的膜。总的来说，我们的工作将SPR的应用扩展到1：1化学计量配体-受体结合的领域之外，进入一个新的巨大的应用领域：溶质如生物分子和药物与脂质的相互作用。
Understanding the interplay between molecules and lipid membranes is fundamental when studying cellular and biotechnological phenomena. Partition between aqueous media and lipid membranes is key to the mechanism of action of many biomolecules and drugs. Quantifying membrane partition, through adequate and robust parameters, is thus essential. Surface Plasmon Resonance (SPR) is a powerful technique for studying 1:1 stoichiometric interactions but has limited application to lipid membrane partition data. We have developed and applied a novel mathematical model for SPR data treatment that enables determination of kinetic and equilibrium partition constants. The method uses two complementary fitting models for association and dissociation sensorgram data. The SPR partition data obtained for the antibody fragment F63, the HIV fusion inhibitor enfuvirtide, and the endogenous drug kyotorphin towards POPC membranes were compared against data from independent techniques. The comprehensive kinetic and partition models were applied to the membrane interaction data of HRC4, a measles virus entry inhibitor peptide, revealing its increased affinity for, and retention in, cholesterol-rich membranes. Overall, our work extends the application of SPR beyond the realm of 1:1 stoichiometric ligand-receptor binding into a new and immense field of applications: the interaction of solutes such as biomolecules and drugs with lipids.